As noted in the subsections of Section
IV-C-1-b-(1), the Director, NIH, may take certain actions with regard to
the NIH Guidelines after the issues have been considered by the
RAC. Some of the actions taken to date
include the following:
Appendix D-1.
Permission is granted to clone foot and mouth disease virus in the EK1
host-vector system consisting of E. coli K-12 and the vector pBR322, all
work to be done at the Plum Island Animal Disease Center.
Appendix D-2.
Certain specified clones derived from segments of the foot and mouth
disease virus may be transferred from Plum Island Animal Disease Center to the facilities
of Genentech, Inc., of South San Francisco, California. Further development of the clones at
Genentech, Inc., has been approved under BL1 + EK1 conditions.
Appendix D-3.
The Rd strain of Hemophilus influenzae can be used as a host for
the propagation of the cloned Tn 10 tet R gene derived from E. coli K-12
employing the non-conjugative Hemophilus plasmid, pRSF0885, under BL1
conditions.
Appendix D-4.
Permission is granted to clone certain subgenomic segments of foot and
mouth disease virus in HV1 Bacillus subtilis and Saccharomyces
cerevisiae host-vector systems under BL1 conditions at Genentech, Inc.,
South San Francisco, California.
Appendix D-5.
Permission is granted to Dr. Ronald Davis of Stanford University to
field test corn plants modified by recombinant DNA techniques under specified
containment conditions.
Appendix D-6.
Permission is granted to clone in E. coli K-12 under BL1 physical
containment conditions subgenomic segments of rift valley fever virus subject
to conditions which have been set forth by the RAC.
Appendix D-7.
Attenuated laboratory strains of Salmonella typhimurium may be
used under BL1 physical containment conditions to screen for the Saccharomyces
cerevisiae pseudouridine synthetase gene.
The plasmid YEp13 will be employed as the vector.
Appendix D-8.
Permission is granted to transfer certain clones of subgenomic segments
of foot and mouth disease virus from Plum Island Animal Disease Center to the
laboratories of Molecular Genetics, Inc., Minnetonka, Minnesota, and to work
with these clones under BL1 containment conditions. Approval is contingent upon review of data on infectivity testing
of the clones by a working group of the RAC.
Appendix D-9.
Permission is granted to Dr. John Sanford of Cornell University to field
test tomato and tobacco plants transformed with bacterial (E.coli K-12)
and yeast DNA using pollen as a vector.
Appendix D-10.
Permission is granted to Drs. Steven Lindow and Nickolas Panopoulos of
the University of California, Berkeley, to release under specified conditions Pseudomonas
syringae, pathovars (pv.) syringae, and Erwinia herbicola
carrying in vitro generated deletions of all or part of the genes
involved in ice nucleation.
Appendix D-11.
Agracetus of Middleton, Wisconsin, may field test under specified
conditions disease resistant tobacco plants prepared by recombinant DNA
techniques.
Appendix D-12.
Eli Lilly and Company of Indianapolis, Indiana, may conduct large-scale
experiments and production involving Cephalosporium acremonium strain
LU4-79-6 under less than Biosafety Level 1 - Large Scale (BL1-LS) conditions.
Appendix D-13.
Drs. W. French Anderson, R. Michael Blaese, and Steven Rosenberg of the
NIH, Bethesda, Maryland, can conduct experiments in which a bacterial gene
coding for neomycin phosphotransferase will be inserted into a portion of the
tumor infiltrating lymphocytes (TIL) of cancer patients using a retroviral
vector, N2. The marked TIL then will be
combined with unmarked TIL, and reinfused into the patients. This experiment is an addition to an ongoing
adoptive immunotherapy protocol in which TIL are isolated from a patient's
tumor, grown in culture in the presence of interleukin-2, and reinfused into
the patient. The marker gene will be
used to detect TIL at various time intervals following reinfusion.
Approval is based on the following four stipulations: (I) there will be no limitation of the
number of patients in the continuing trial; (ii) the patients selected will
have a life expectancy of about 90 days; (iii) the patients give fully informed
consent to participate in the trial; and (iv) the investigators will provide
additional data before inserting a gene for therapeutic purposes. (Protocol
#8810-001)
Appendix D-14.
U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID)
may conduct certain experiments involving products of a yellow fever virus
originating from the 17-D yellow fever clone at the Biosafety Level 3
containment level using HEPA filters and vaccination of laboratory personnel.
In addition, USAMRIID may conduct certain experiments
involving vaccine studies of Venezuelan equine encephalitis virus at the
Biosafety Level 3 containment level using HEPA filters and vaccination of
laboratory personnel.
Appendix D-15.
Drs. R. Michael Blaese and W. French Anderson of the NIH, Bethesda,
Maryland, can conduct experiments in which a gene coding for adenosine
deaminase (ADA) will be inserted into T lymphocytes of patients with severe combined
immunodeficiency disease, using a retroviral vector, LASN. Following insertion of the gene, these T
lymphocytes will be reinfused into the patients. The patients will then be followed for evidence of clinical
improvement in the disease state, and measurement of multiple parameters of
immune function by laboratory testing.
Approval is based on the following two stipulations: (I) that intraperitoneal administration of
transduced T lymphocytes not be used before clearance by the Chair of the Recombinant
DNA Advisory Committee; and (ii) that the number of research patients be
limited to 10 at this time.
In addition to the conditions outlined in the initial
approval, patients may be given a supplement of a CD-34+-enriched peripheral
blood lymphocytes (PBL) which have been placed in culture conditions that favor
progenitor cell growth. This enriched
population of cells will be transduced with the retroviral vector,
G1NaSvAd. G1NaSvAd is similar to LASN,
yet distinguishable by PCR. LASN has
been used to transduce peripheral blood T lymphocytes with the ADA gene. Lymphocytes and myeloid cells will be
isolated from patients over time and assayed for the presence of the LASN or
G1NaSvAd vectors. The primary
objectives of this protocol are to transduce CD 34+ peripheral blood cells with
the adenosine deaminase gene, administer these cells to patients, and determine
if such cells can differentiate into lymphoid and myeloid cells in vivo. There is a potential for benefit to the
patients in that these hematopoietic progenitor cells may survive longer, and
divide to yield a broader range of gene-corrected cells. (Protocol #9007-002)
Appendix D-16.
Dr. Steven A. Rosenberg of the National Institutes of Health, Bethesda,
Maryland, can conduct experiments on patients with advanced melanoma who have
failed all effective therapy. These
patients will be treated with escalating doses of autologous tumor infiltrating
lymphocytes (TIL) transduced with a gene coding for tumor necrosis factor
(TNF). Escalating numbers of transduced
TIL will be administered at three weekly intervals along with the
administration of interleukin-2 (IL-2).
The objective is to evaluate the toxicity and possible therapeutic
efficacy of the administration of tumor infiltrating lymphocytes (TIL)
transduced with the gene coding for TNF. (Protocol #9007-003)
Appendix D-17.
Dr. Malcolm K. Brenner of St. Jude Children's Research Hospital of
Memphis, Tennessee, can conduct experiments on patients with acute myelogenous
leukemia (AML). Using the LNL6
retroviral vector, the autologous bone marrow cells will be transduced with the
gene coding for neomycin resistance.
The purpose of this gene marking experiment is to determine whether the
source of relapse after autologous bone marrow transplantation for acute
myelogenous leukemia is residual malignant cells in the harvested marrow or
reoccurrence of tumor in the patient.
Determining the source of relapse should indicate whether or not purging
of the bone marrow is a necessary procedure. (Protocol #9102-004)
Appendix D-18.
Dr. Malcolm K. Brenner of St. Jude Children's Research Hospital of
Memphis, Tennessee, can conduct experiments on pediatric patients with Stage D
(disseminated) neuroblastoma who are being treated with high-dose carboplatin
and etoposide in either phase I/II or phase II trials. All the patients in these studies will be
subjected to bone marrow transplantation since it will allow them to be exposed
to chemoradiation that would be lethal were it not for the availability of
stored autologous marrow for rescue.
The bone marrow cells of these patients will be transduced with the gene
coding for neomycin resistance using the LNL6 vector. The purpose of this gene marking study is to determine whether
the source of relapse after autologous bone marrow transplantation is residual
malignant cells in the harvested marrow or residual disease in the
patient. Secondly, it is hoped to
determine the contribution of marrow autographs to autologous reconstitution.
(Protocol #9105-005/9105-006)
Appendix D-19.
Dr. Albert B. Deisseroth of the MD Anderson Cancer Center of Houston,
Texas, can conduct experiments on patients with chronic myelogenous leukemia
who have been reinduced into a second chronic phase or blast cells. The patients in these studies will receive
autologous bone marrow transplantation.
Using the LNL6 vector, the bone marrow cells will be transduced with the
gene coding for neomycin resistance.
The purpose of these marking studies is to determine if the origin of
relapse arises from residual leukemic cells in the patients or from viable
leukemic cells remaining in the bone marrow used for autologous
transplantation. (Protocol #9105-007)
Appendix D-20.
Drs. Fred D. Ledley and Savio L. C. Woo of Baylor College of Medicine of
Houston, Texas, can conduct experiments on pediatric patients with acute
hepatic failure who are identified as candidates for hepatocellular
transplantation. Using the LNL6 vector,
the hepatocytes will be transduced with the gene coding for neomycin resistance. The purpose of using a genetic marker is to
demonstrate the pattern of engraftment of transplanted hepatocytes and to help
determine the success or failure of engraftment. (Protocol #9105-008)
Appendix D-21.
Dr. Steven A. Rosenberg of the National Institutes of Health, Bethesda,
Maryland, can conduct experiments on patients with advanced melanoma, renal
cell cancer, and colon carcinoma who have failed all effective therapy. In an attempt to increase these patients'
immune responses to the tumor, the tumor necrosis factor gene or the
interleukin-2 gene will be introduced into a tumor cell line established from
the patient. These gene-modified
autologous tumor cells will then be injected into the thigh of the patient. To further utilize the immune system of the
patient to fight the tumor, stimulated lymphocytes will be cultured from either
the draining regional lymph nodes or the injected tumor itself. The patients will be evaluated for antitumor
effects engendered by the injection of the gene modified tumor cells themselves
as well as after the infusion of the cultured lymphocytes. (Protocol
#9110-010/9110-011)
Appendix D-22.
Dr. James M. Wilson of the University of Michigan Medical Center of Ann
Arbor, Michigan, can conduct experiments on three patients with the homozygous
form of familial hypercholesterolemia.
Both children and adults will be eligible for this therapy. In an attempt to correct the basic genetic
defect in this disease, the gene coding for the low-density lipoprotein (LDL)
receptor will be introduced into liver cells taken from the patient. The gene-corrected hepatocytes will then be
infused into the portal circulation of the patient through an indwelling
catheter. The patients will be
evaluated for engraftment of the these treated hepatocytes through a series of
metabolic studies; three months after gene therapy, a liver biopsy will be
taken and analyzed for the presence of recombinant derived RNA and DNA to
document the presence of the gene coding for the normal LDL receptor. (Protocol
#9110-012)
Appendix D-23.
Dr. Michael T. Lotze of the University of Pittsburgh School of Medicine,
Pittsburgh, Pennsylvania, can conduct experiments on 20 patients with
metastatic melanoma who have failed conventional therapy. A gene transfer experiment will be
performed, transducing the patients' tumor infiltrating lymphocytes (TILs) with
the gene for neomycin resistance.
Through the use of this gene marking technique, it is proposed to
determine how long TIL cells can be detected in vivo in the peripheral
blood of the patients, and how the administration of interleukin-2 and
interleukin-4 affects localization and survival of TIL cells in tumor sites.
(Protocol #9105-009)
Appendix D-24.
Dr. Gary J. Nabel of the University of Michigan Medical School, Ann
Arbor, Michigan, can conduct gene therapy experiments on twelve patients with
melanoma or adenocarcinoma. Patient
population will be limited to adults over the age of 18 and female patients
must be postmenopausal or have undergone tubal ligation or orchiectomy. The patient's immune response will be
stimulated by the introduction of a gene encoding for a Class I MHC protein,
HLA-B7, in order to enhance tumor regression.
DNA/liposome-mediated transfection techniques will be used to directly
transfer this foreign gene into tumor cells.
HLA-B7 expression will be confirmed in vivo, and the immune
response stimulated by the expression of this antigen will be
characterized. These experiments will
be analyzed for their efficacy in treating cancer. (Protocol #9202-013)
Appendix D-25.
Kenneth Cornetta of Indiana University, Indianapolis, Indiana, can
conduct gene transfer experiments on up to 10 patients with acute myelogenous
leukemia (AML) and up to 10 patients with acute lymphocytic leukemia (ALL). The patient population will be limited to
persons between 18 and 65 years of age.
Using the LNL-6 vector, autologous bone marrow cells will be marked with
the neomycin resistance gene. Gene
marked and untreated bone marrow cells will be reinfused at the time of bone
marrow transplantation. Patients will
then be monitored for evidence of the neomycin resistance gene in peripheral
blood and bone marrow cells in order to determine whether relapse of their
disease is a result of residual malignant cells remaining in the harvested
marrow or inadequate ablation of the tumor cells by chemotherapeutic
agents. Determining the source of
relapse may indicate whether or not purging of the bone marrow is a necessary
procedure for these leukemia patients.
Further studies will be performed in order to determine the percentage
of leukemic cells that contain the LNL-6 vector and the clonality of the marked
cells. (Protocol #9202-014)
Appendix D-26.
Dr. James S. Economou of the University of California, Los Angeles, can
conduct gene transfer experiments on 20 patients with metastatic melanoma and
20 patients with renal cell carcinoma.
These patients will be treated with various combinations of
tumor-infiltrating lymphocytes and peripheral blood leukocytes, including CD8
and CD4 subsets of both types of cells.
These effector cell populations will be given in combination with
interleukin-2 (IL-2) in the melanoma patients and IL-2 plus alpha interferon in
the renal cell carcinoma patients. The
effector cells will be transduced with the neomycin resistance gene using
either the LNL6 or G1N retroviral vectors.
This "genetic marking" of the tumor-infiltrating lymphocytes
and peripheral blood lymphocytes is designed to answer questions about the
trafficking of these cells, their localization to tumors, and their in vivo
life span. (Protocol #9202-015)
Appendix D-27.
Drs. Philip Greenberg and Stanley R. Riddell of the Fred Hutchinson
Cancer Research Center, Seattle, Washington, may conduct gene transfer experiments
on 15 human immunodeficiency virus (HIV) seropositive patients (18-45 years
old) undergoing allogeneic bone marrow transplantation for non-Hodgkin's
lymphoma and 15 HIV-seropositive patients (18-50 years old) who do not have
acquired immunodeficiency syndrome (AIDS)-related lymphoma and who are not
undergoing bone marrow transplantation to evaluate the safety and efficacy of
HIV-specific cytotoxic T lymphocyte (CTL) therapy. CTL will be transduced with a retroviral vector (HyTK) encoding a
gene that is a fusion product of the hygromycin phosphotransferase gene (HPH)
and the herpes simplex virus thymidine kinase (HSV-TK) gene. This vector will deliver both a marker gene
and an ablatable gene in these T cell clones in the event that patients develop
side effects as a consequence of CTL therapy.
Data will be correlated over time, looking at multiple parameters of HIV
disease activity. The objectives of
these studies include evaluating the safety and toxicity of CTL therapy,
determining the duration of in vivo survival of HIV-specific CTL clones,
and determining if ganciclovir therapy can eradicate genetically modified,
adoptively transferred CTL cells. (Protocol #9202-017)
Appendix D-28.
Dr. Malcolm Brenner of St. Jude Children's Research Hospital, Memphis,
Tennessee, can conduct gene therapy experiments on twelve patients with
relapsed/refractory neuroblastoma who have relapsed after receiving autologous
bone marrow transplant. In an attempt
to stimulate the patient's immune response, the gene coding for Interleukin-2
(IL-2) will be used to transduce tumor cells, and these gene-modified cells
will be injected subcutaneously in a Phase 1 dose escalation trial. Patients will be evaluated for an anti-tumor
response. (Protocol #9206-018)
Appendix D-29.
Drs. Edward Oldfield, Kenneth Culver, Zvi Ram, and R. Michael Blaese of
the National Institutes of Health, Bethesda, Maryland, can conduct gene therapy
experiments on ten patients with primary malignant brain tumors and ten
patients with lung cancer, breast cancer, malignant melanoma, or renal cell
carcinoma who have brain metastases.
The patient population will be limited to adults over the age of 18.
Patients will be divided into two groups based on the
surgical accessibility of their lesions.
Both surgically accessible and surgically inaccessible lesions will
receive intra-tumoral injections of the retroviral Herpes simplex thymidine
kinase (HS-tk) vector-producer cell line, G1TkSvNa, using a guided stereotaxic
approach. Surgically accessible lesions
will be excised seven days after stereotaxic injection, and the tumor bed will
be infiltrated with the HS-tk producer cells.
The removed tumor will be evaluated for the efficiency of transduction. Ganciclovir (GCV) will be administered
beginning on the fifth postoperative day.
In the case of surgically inaccessible lesions, the patients will
receive intravenous therapy with GCV seven days after receiving the
intra-tumoral injections of the retroviral HS-tk vector-producer cells.
(Protocol #9206-019)
Appendix D-30.
Dr. Albert D. Deisseroth of MD Anderson Cancer Center, Houston, Texas,
can conduct gene transfer experiments on ten patients who have developed blast
crisis or accelerated phase chronic myelogenous leukemia (CML). The retroviral vectors G1N and LNL6 which
code for neomycin resistance will be used to transduce autologous peripheral
blood and bone marrow cells that have been removed and stored at the time of
cytogenetic remission or re-induction of chronic phase in Philadelphia chromosome
positive CML patients. Following
reinduction of the chronic phase of CML and preparative chemotherapy, patients
will be infused with the transduced autologous cells.
This protocol is designed to determine the cause of relapse
of CML. If polyclonal CML neomycin
marked blastic cells appear at the time of relapse, their presence will
indicate that relapse arises from the leukemic CML blast cells present in the
autologous cells infused following chemotherapy. If residual systemic disease contributes to relapse, the neomycin
resistance gene will not be detected in the CML leukemic blasts at the time of
relapse.
This study will compare the relative contributions of the
peripheral blood and bone marrow to generate hematopoietic recovery after bone marrow
transplantation and evaluate purging and selection of peripheral blood or bone
marrow as a source of stem cells for transplant. The percentage of neomycin resistant CML cells which are leukemic
will be determined by PCR analysis and detection of bcr-abl mRNA. (Protocol
#9206-020)
Appendix D-31.
Dr. Cynthia Dunbar of the National Institutes of Health, Bethesda,
Maryland, can conduct gene transfer experiments on up to 48 patients with
multiple myeloma, breast cancer, or chronic myelogenous leukemia. The retroviral vectors G1N and LNL6 will be
used to transfer the neomycin resistance marker gene into autologous bone
marrow and peripheral blood stem cells in the presence of growth factors to
examine hematopoietic reconstitution after bone marrow transplantation. The efficiency of transduction of both short
and long term autologous bone marrow reconstituting cells will be examined.
Autologous bone marrow and CD34+ peripheral blood stem cells
will be enriched prior to transduction.
Myeloma and CML patients will receive both autologous bone marrow and
peripheral blood stem cell transplantation.
These separate populations will be marked with both the G1N and LNL6
retroviral vectors. If short and long
term marking experiments are successful, important information may be obtained
regarding the biology of autologous reconstitution, the feasibility of
retroviral gene transfer into hematopoietic cells, and the contribution of
viable tumor cells within the autograft to disease relapse. (Protocol
#9206-023/9206-024/9206-025)
Appendix D-32.
Dr. Bernd Gansbacher of the Memorial Sloan-Kettering Cancer Center, New
York, New York, can conduct gene therapy experiments on twelve patients over 18
years of age with metastatic melanoma who are HLA-A2 positive and who have
failed conventional therapy. This is a
phase I study to examine whether allogeneic HLA-A2 matched melanoma cells
expressing recombinant human Interleukin-2 (IL-2) can be injected
subcutaneously and used to create a potent tumor specific immune response
without producing toxicity. By allowing
the tumor cells to present the MHC Class I molecule as well as the secreted
IL-2, a clonal expansion of tumor specific effector cells is expected. These effector populations may access
residual tumor at distant sites via the systemic circulation. (Protocol
#9206-021)
Appendix D-33.
Dr. Bernd Gansbacher of the Memorial Sloan-Kettering Cancer Center, New
York, New York, can conduct gene therapy experiments on twelve patients over 18
years of age with renal cell carcinoma who are HLA-A2 positive and who have
failed conventional therapy. This Phase
I study will examine whether allogeneic HLA-A2 matched renal cell carcinoma
cells expressing recombinant human Interleukin-2 (IL-2) can be injected
subcutaneously and used to create a potent tumor specific immune response
without producing toxicity. By allowing
the tumor cells to present the MHC Class I molecule as well as the secreted
IL-2, a clonal expansion of tumor specific effector cells is expected. These effector populations may access
residual tumor at distant sites via the systemic circulation. (Protocol
#9206-022)
Appendix D-34.
Dr. Michael T. Lotze, University of Pittsburgh, Pittsburgh,
Pennsylvania, can conduct experiments on twenty patients with metastatic,
and/or unresectable, locally advanced melanoma, renal cell carcinoma, breast
cancer, or colon cancer who have failed standard therapy. Patients will receive multiple subcutaneous
injections of autologous tumor cells combined with an autologous fibroblast
cell line that has been transduced in vitro with the gene coding for
Interleukin-4 (IL-4) to augment the in vivo antitumor effect. Patients will be monitored for antitumor
effect by PCR analysis and multiple biopsy of the injection site. (Protocol #9209-033)
Appendix D-35.
Dr. Friedrich G. Schuening, Fred Hutchinson Cancer Research Center,
Seattle, Washington, can conduct human gene transfer experiments on patients ≥ 18
years of age with breast cancer, Hodgkin's disease, or non-Hodgkin's
lymphoma. A total of 10 patients per
year will be enrolled in the studies over a period of four years. Patients will undergo autologous bone marrow
transplantation with a selected population of Interleukin-3 (IL-3) or granulocyte
colony-stimulating factor (G-CSF) stimulated CD34(+) peripheral blood
repopulating cells (PBRC) that have been transduced with the gene coding for
neomycin resistance (neoR) using the retroviral vector, LN. Patients will be continuously monitored for
neoR to determine the relative contribution of autologous PBRCs to
long-term hematopoietic reconstitution.
Demonstration of long-term contribution of autologous PBRC to
hematopoiesis will enable the use of PBRC alone for autologous transplants and
suggest the use of PBRC as long-term carriers of therapeutically relevant
genes. (Protocol #9209-027/9209-028)
Appendix D-36.
Dr. Friedrich G. Schuening, Fred Hutchinson Cancer Research Center,
Seattle, Washington, can conduct human gene transfer experiments on patients ≥ 18
years of age with breast cancer, Hodgkin's disease, or non-Hodgkin's
lymphoma. A total of 5 patients per
year will be enrolled in the study over a period of four years. Patients will undergo allogeneic bone marrow
transplant with granulocyte colony-stimulating factor (G-CSF) stimulated
CD34(+) PBRC harvested from an identical twin that have been transduced with
neoR using the retroviral vector, LN. Patients will be continuously monitored for neoR to
determine the relative contribution of G-CSF stimulated allogeneic PBRCs to
long-term bone marrow engraftment.
Demonstration of long-term contribution of allogeneic PBRC to
hematopoiesis will enable the use of PBRC alone for allogeneic transplants and
suggest the use of PBRC as long-term carriers of therapeutically relevant
genes. (Protocol #9209-029)
Appendix D-37.
Dr. Malcolm K. Brenner of St. Jude Children's Hospital, Memphis,
Tennessee, and Dr. Bonnie J. Mills of Baxter Healthcare Corporation, Santa Ana,
California, can conduct a multicenter uncontrolled human gene transfer experiment
on 12 patients ≤ 21 years of age with Stage D Neuroblastoma in first or
second marrow remission. Autologous
bone marrow cells will be separated into two fractions, purged and
unpurged. Each fraction will be
transduced with the neoR gene by either LNL6 or G1Na. Patients will be monitored by the polymerase
chain reaction (PCR) for the presence of neoR. The protocol is designed to evaluate the
safety and efficacy of the Neuroblastoma Bone Marrow Purging System following
high dose chemotherapy. (Protocol #9209-032)
Appendix D-38.
Drs. Carolyn Keierleber and Ann Progulske-Fox of the University of
Florida, Gainesville, Florida, can conduct experiments involving the
introduction of a gene coding for tetracycline resistance into Porphyromonas
gingivalis at a physical containment level of Biosafety Level-2 (BL-2).
Appendix D-39.
Dr. Scott M. Freeman of Tulane University Medical Center, New Orleans,
Louisiana, can conduct experiments on patients with epithelial ovarian
carcinoma who have clinical evidence of recurrent, progressive, or residual
disease who have no other therapy available to prolong survival. Patients will be injected intraperitoneally
with the irradiated PA-1 ovarian carcinoma cell line which has been transduced
with the herpes simplex thymidine kinase (HSV-TK) gene. The patients will then receive ganciclovir
therapy. Previous, data indicates that
HSV-TK+ tumor cells exhibit a killing effect on HSV-TK- cells when exposed to
ganciclovir therapy. Patients will be
evaluated for safety and side effects of this treatment. (Protocol #9206-016)
Appendix D-40.
Dr. Michael J. Welsh, Howard Hughes Medical Institute Research
Laboratories, University of Iowa College of Medicine, Iowa City, Iowa, may
conduct experiments on 3 cystic fibrosis (CF) patients ≥ 18 years of age with mild to
moderate disease. This Phase I study
will determine the: (1) in vivo
safety and efficacy of the administration of the replication-deficient type 2
adenovirus vector, Ad2/CFTR-1, to the nasal epithelium; (2) efficacy in
correcting the CF chloride transport defect in vivo; and (3) effect of
adenovirus vector dosage on safety and efficacy. (Protocol #9212-036)
Appendix D-41.
Dr. Ronald G. Crystal, National Institutes of Health, Bethesda,
Maryland, may conduct experiments on 10 cystic fibrosis (CF) patients ≥ 21
years of age. Patients will receive an
initial administration of the replication-deficient type 5 adenovirus vector,
AdCFTR, to their left nares. If no
toxicity is observed from intranasal administration, patients will receive a
single administration of AdCFTR to the respiratory epithelium of their left
large bronchi. Five groups of patients
(2 patients per group) will be studied based on increased dosage administration
of AdCFTR. This study will determine
the: (1) in vivo safety and
efficacy of the administration of AdCFTR into the respiratory epithelium; (2)
efficacy of the correction of the biologic abnormalities of CF in the
respiratory epithelium; (3) duration of
the biologic correction; (4) efficacy of the correction of the abnormal
electrical potential difference of the airway epithelial sheet; (5) clinical
parameters relevant to the disease process; and (6) if humoral immunity
develops against AdCFTR sufficient to prevent repeat administration. (Protocol
#9212-034)
Appendix D-42.
Dr. Kenneth Culver, Iowa Methodist Medical Center, Des Moines, Iowa, and
Dr. John Van Gilder, University of Iowa, Iowa City, Iowa, may conduct
experiments on 15 patients ≥ 18 years of age with recurrent malignant primary brain
tumors or lung, melanoma, renal cell carcinoma, or breast carcinoma brain
metastases who have failed standard therapy for their disease. Patient eligibility will be limited to those
patients who have measurable residual tumor immediately following the
post-operative procedure as demonstrated by imaging studies. The number of patients treated will be
equally divided between the Iowa Methodist Medical Center and the University of
Iowa. If a positive response is
observed in any of the first 15 patients, the investigators may submit a
request to treat an additional 15 patients.
Following surgical debulking, patients will receive a
maximum of 3 intralesional injections of the G1TkSvNa vector- producing cell
line (VPC) to induce regression of residual tumor cells by ganciclovir (GCV)
therapy. Patients who demonstrate
stable disease for a minimum of 6 months following this treatment will be
eligible for additional VPC injections and subsequent GCV therapy. (Protocol
#9303-037)
Appendix D-43.
Drs. Malcolm Brenner, Robert Krance, Helen E. Heslop, Victor Santana,
and James Ihle, St. Jude Children's Research Hospital, Memphis, Tennessee, may
conduct experiments on 35 patients ≥ 1 year and ≤ 21 years of age at the time
of initial diagnosis of acute myelogenous leukemia (AML). The investigators will use the two
retroviral vectors, LNL6 and G1Na, to determine the efficacy of the bone marrow
purging techniques: 4-hydroxyperoxicyclophosphamide
and interleukin-2 (IL-2) activation of endogenous cytotoxic effector cells, in
preventing relapse from the reinfusion of autologous bone marrow cells.
(Protocol #9303-039)
Appendix D-44.
Drs. Helen E. Heslop, Malcolm Brenner, and Cliona Rooney, St Jude
Children's Research Hospital, Memphis, Tennessee, may conduct experiments of 35
patients ≤
21 years of age who will be recipients of mismatched-related or phenotypically
similar unrelated donor marrow grafts for leukemia. In this Phase I dose escalation study, spontaneous lymphoblastoid
cell lines will be established that express the same range of Epstein-Barr
Virus (EBV) encoded proteins as the recipient.
These EBV-specific cell lines will be transduced with LNL6 or G1Na and
readministered at the time of bone marrow transplant. This study will determine:
(1) survival and expansion of these EBV-specific cell lines in vivo,
(2) the ability of these adoptively transferred cells to confer protection
against EBV infection, and (3) appropriate dosage and administration schedules.
(Protocol #9303-038)
Appendix D-45. Drs. Robert W. Wilmott and Jeffrey Whitsett, Children's Hospital
Medical Center, Cincinnati, Ohio, and Dr. Bruce Trapnell, Genetic Therapy,
Inc., Gaithersburg, Maryland, may conduct experiments on 15 cystic fibrosis
(CF) patients who have mild to moderate disease ≥ 21 years of age. The replication-deficient type 5 adenovirus
vector, Av1CF2, will be administered to the nasal and lobar bronchial
respiratory tract of patients. This
study will demonstrate the: (1)
expression of normal cystic fibrosis transmembrane conductance regulator (CFTR)
mRNA in vivo, (2) synthesis of CFTR protein, and (3) correction of
epithelial cell cAMP dependent Cl- permeability. The pharmacokinetics of CFTR expression and
ability to re-infect the respiratory tract with AvCF2 will be determined. Systemic and local immunologic consequences
of Av1CF2 infection, the time of viral survival, and potential for
recombination or complementation of the virus will be monitored. (Protocol
#9303-041)
Appendix D-46.
Dr. James M. Wilson of the University of Pennsylvania Medical Center,
Philadelphia, Pennsylvania, may conduct experiments on 20 adult patients with
advanced cystic fibrosis lung disease.
An isolated segment of the patients' lung will be transduced with the E1
deleted, replication-incompetent adenovirus vector, Ad.CB-CFTR using a bronchoscope for gene delivery. Ad.CB-CFTR contains the human gene encoding
the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Pulmonary biopsies will be obtained by
bronchoscopy at 4 days, 6 weeks, and 3 months following treatment. Patients will be monitored for evidence of
CFTR gene transfer and expression, immunological responses to CFTR or
adenovirus proteins, and toxicity. (Protocol #9212-035)
Appendix D-47.
Dr. Hilliard F. Seigler of Duke University Medical Center, Durham, North
Carolina, may conduct experiments on 20 patients with disseminated malignant
melanoma. Autologous tumor cells will
be transduced with a retroviral vector, pHuγ-IFN, that contains the gene encoding human γ-IFN. Following lethal irradiation, the transduced cells will be
readministered to patients for the purpose of generating cytotoxic T cells that
are tumor specific along with the up-regulation of Class I major histocompatibility
antigens. Patients will be monitored
for clinical regression of tumors and generation of tumor-specific cytotoxic T
lymphocytes. (Protocol #9306-043)
Appendix D-48.
Drs. Stefan Karlsson and Cynthia Dunbar of the National Institutes of Health,
Bethesda, Maryland, and Dr. Donald B. Kohn of the Children’s Hospital of Los
Angeles, Los Angeles, California, may conduct experiments on 10 patients with
Gaucher disease. CD34(+) hematopoietic
stem cells will be isolated from bone marrow or from peripheral blood treated
with granulocyte-colony stimulating factor.
CD34(+) cells will be transduced with a retrovirus vector, G1Gc,
containing cDNA encoding human glucocerebrosidase and administered
intravenously. Patients will be
monitored for toxicity and glucocerebrosidase expression. (Protocol #9306-047)
Appendix D-49.
Dr. Gary J. Nabel of the University of Michigan Medical Center, Ann
Arbor, Michigan, may conduct experiments on 12 patients with AIDS to be divided
into 4 experimental groups. CD4(+)
lymphocytes will be isolated from peripheral blood and transduced with Rev M10,
a transdominant inhibitory mutant of the rev gene of the human
immunodeficiency virus (HIV).
Transduction of the rev mutant will be mediated either by the
retrovirus vector, PLJ-cREV M10, or by particle-mediated gene transfer of
plasmid DNA. Patients will be monitored
for survival of the transduced CD4(+) cells by polymerase chain reaction and
whether Rev M10 can confer protection against HIV infection to CD4(+) cells.
(Protocol #9306-049)
Appendix D-50.
Dr. Gary J. Nabel of the University of Michigan Medical Center, Ann
Arbor, Michigan, may conduct experiments on 24 patients with advanced
cancer. Patients will undergo in
vivo transduction with DNA/liposome complexes containing genes encoding the
HLA-B7 histocompatibility antigen and beta-2 microglobulin in a non-viral
plasmid. These DNA/liposome complexes
will be administered either by intratumoral injection or catheter delivery. Patients will be monitored for enhanced immune
responses against tumor cells, and safe and effective doses will be determined.
(Protocol #9306-045)
Appendix D-51.
Dr. John A. Barranger of the University of Pittsburgh, Pittsburgh,
Pennsylvania, may conduct experiments on 5 patients with Gaucher disease. The CD34(+) hematopoietic stem cells will be
isolated from peripheral blood and transduced in vitro with the
retrovirus vector, N2-Sv-GC, encoding the glucocerebrosidase (GC) enzyme. Following reinfusion of the transduced
cells, patients will be monitored by PCR analysis for GC expression in
peripheral blood leukocytes. Patients
currently receiving GC replacement therapy and who demonstrate clinical
responsiveness will be withdrawn from exogenous GC therapy. Patients not previously treated with
exogenous GC, will be monitored for clinical reversal of lipid storage
symptoms. (Protocol #9306-046)
Appendix D-52.
Dr. Robert Walker of the National Institutes of Health, Bethesda,
Maryland, may conduct experiments on 12 HIV-infected patients who have a
seronegative identical twin. CD4(+) and
CD8(+) cells will be isolated from the seronegative twin and induced to
polyclonal proliferation with anti-CD3 and interleukin-2. The enriched population of cells will be
transduced with either LNL6 or G1Na, which contain the neoR
gene. The transduced cells will be
expanded in tissue culture and administered to the HIV-infected twin. Patients will be monitored for immune
function and the presence of marked cells. (Protocol #9209-026)
Appendix D-53.
Dr. Corey Raffel of the Children’s Hospital Los Angeles, Los Angeles,
California, and Dr. Kenneth Culver of Iowa Methodist Medical Center, Des
Moines, Iowa, may conduct experiments on 30 patients between 2 and 18 years of
age with recurrent malignant astrocytoma.
Fifteen patients will be accrued into this study initially. If at least one patient responds to therapy,
an additional 14 patients will be treated.
Patients with either surgically accessible or non-accessible tumors will
be treated with the vector producing cell line (PA317) carrying the retrovirus
vector, G1TkSvNa. This vector will
transduce tumor cells in vivo with the Herpes simplex thymidine
kinase (HS-tk) gene that renders the cells sensitive to killing by
ganciclovir. Surgically accessible patients
will undergo surgical debulking of their tumor followed by repeated
administration of the HS-tk vector producer cells into the tumor bed. Children with unresectable tumors will
undergo stereotaxic injection of vector producer cells into tumors. (Protocol
#9306-050)
Appendix D-54.
Dr. Jeffrey E. Galpin of the University of Southern California, Los
Angeles, California, and Dr. Dennis A. Casciato of the University of
California, Los Angeles, California, may conduct experiments on 15 HIV(+) asymptomatic
patients. Patients will receive 3
monthly intramuscular injections of the retrovirus vector (N2IIIBenv) encoding
the HIV-1 IIIB envelope protein.
Patients will be monitored for acute toxicity, CD4 levels, HIV-specific
CTL responses, and viral burdens. (Protocol #9306-048)
Appendix D-55.
Drs. Charles Hesdorffer and Karen Antman of Columbia University College
of Physicians and Surgeons, New York, New York, may conduct experiments on 20
patients with advanced breast, ovary, and brain cancer. CD34(+) hematopoietic stem cells will be
isolated from bone marrow, transduced with the retrovirus vector, PHaMDR1/A,
and readministered to patients.
Patients will be monitored for the presence and expression of the MDR-1
gene. The investigators will determine
whether MDR-1 expression increases following chemotherapy. (Protocol #9306-051)
Appendix D-56.
Dr. Enzo Paoletti of Virogenetics Corporation, Troy, New York, may
conduct experiments with poxvirus vectors NYVAC, ALVAC, and TROVAC at Biosafety
Level 1.
Appendix D-57.
Drs. Richard C. Boucher and Michael R. Knowles of the University of
North Carolina, Chapel Hill, North Carolina, may conduct experiments on 9
patients (18 years old or greater) with cystic fibrosis to test for the safety
and efficacy of an E1-deleted recombinant adenovirus containing the cystic
fibrosis transmembrane conductance regulator (CFTR) cDNA, Ad.CB-CFTR. A single dose of 108, 3 x 109
or 1011 pfu/ml will be administered to the nasal cavity of 3
patients in each dose group. Patients
will be monitored by nasal lavage and biopsy to assess safety and restoration
of normal epithelial function. (Protocol #9303-042)
Appendix D-58.
Dr. Joyce A. O'Shaughnessy of the National Institutes of Health,
Bethesda, Maryland, may conduct experiments on 18 patients (18-60 years old)
with Stage IV breast cancer who have achieved a partial or complete response to
induction chemotherapy. This study will
determine the feasibility of obtaining engraftment of CD34(+) hematopoietic
stem cells transduced by a retroviral vector, G1MD, and expressing a cDNA for
the human multi-drug resistance-1 (MDR-1) gene following high dose
chemotherapy, and whether the transduced MDR-1 gene confers drug resistance to
hematopoietic cells and functions as an in vivo dominant selectable
marker. Patients will be monitored for
evidence of myeloprotection and presence of the transduced MDR-1 gene."
(Protocol #9309-054)
Appendix D-59.
Drs. Larry E. Kun, R. A. Sanford, Malcolm Brenner, and Richard L.
Heideman of St. Jude Children's Research Hospital, Memphis, Tennessee, and Dr.
Edward H. Oldfield of the National Institutes of Health, Bethesda, Maryland,
may conduct experiments on 6 patients (3-21 years old) with progressive or
recurrent malignant supratentorial tumors resistant to standard therapies. Mouse cells producing the retroviral vector
containing the herpes simplex thymidine kinase
gene (G1TKSVNa) will be instilled into the tumor areas via multiple
stereotactically placed cannulas.
Patients will be treated with ganciclovir to eliminate cells expressing
the transduced gene. Patients will be
monitored for central nervous system, hematologic, renal or other toxicities,
and for anti-tumor responses by magnetic resonance imaging studies. (Protocol
#9309-055)
Appendix D-60.
The physical containment level may be reduced from Biosafety Level 3 to
Biosafety Level 2 for a Semliki Forest Virus (SFV) vector expression system of
Life Technologies, Inc., Gaithersburg, Maryland.
Appendix D-61.
Dr. Albert B. Deisseroth of the University of Texas MD Anderson Cancer
Center, Houston, Texas, may conduct experiments on 10 patients (≥ 16
to ≤ 60
years of age) with chronic lymphocytic leukemia. Autologous peripheral blood and bone marrow cells will be removed
from patients following chemotherapy and marked by transduction with two
distinguishable retroviral vectors, G1Na and LNL6, containing the neomycin
resistance gene. The gene marked cells
will be reinfused into patients to determine the efficiency of bone marrow
purging and the origin of relapse following autologous bone marrow
transplantation. (Protocol #9209-030)
Appendix D-62.
Dr. Jonathan Simons of the Johns Hopkins Oncology Center, Baltimore,
Maryland, may conduct experiments on 26 patients (≥ 18 years of age) with
metastatic renal cell carcinoma to evaluate the safety and tolerability of
intradermally injected autologous irradiated tumor cells transduced with the
retrovirus vector, MFG, containing the human granulocyte-macrophage colony
stimulating factor gene. Acute and
long-term clinical toxicities and in vitro and in vivo induction
of specific anti-tumor immune responses will be monitored. (Protocol #9303-040)
Appendix D-63.
Dr. Albert B. Deisseroth of the University of Texas MD Anderson Cancer
Center, Houston, Texas, may conduct experiments on 20 patients (≥ 18
and ≤ 60
years old) with ovarian cancer. A
murine viral vector was constructed from the third generation of L series
retroviruses with the insert of the human multi-drug resistance-1 (MDR-1)
transduced gene. The investigators will
assess the safety and feasibility of administering CD34 (+) autologous
peripheral blood and bone marrow cells.
Patients will be monitored for the presence of the MDR-1 gene and for
the effect of gene transfer on hematopoietic function following the
transplantation. (Protocol #9306-044)
Appendix D-64.
Dr. Joseph Ilan of the Case Western Reserve University School of
Medicine and University Hospital of Cleveland, Cleveland, Ohio, may conduct
experiments on 12 patients (≥ 18 years of age) with advanced brain cancer. Human malignant glioma tumor cells will be
cultured, transfected with Epstein-Barr virus (EBV)-based vector, anti-Insulin
growth factor-I, lethally irradiated, and injected subcutaneously into patients
in an attempt to express antisense Insulin growth factor-1. Patients will be monitored for toxicity and
immunologic responses to the vector. (Protocol #9306-052)
Appendix D-65.
Drs. James S. Economou and John Glaspy of the University of California,
Los Angeles, California, may conduct experiments on 30 patients (≥ 18
to ≤ 70
years of age) with metastatic melanoma.
A human melanoma cell line (M-24) will be transduced with the retroviral
vector, G1NaCvi2, expressing the human interleukin-2 (IL-2) gene. The IL-2 producing cells will be mixed with
the patient's autologous tumor cells, irradiated, and injected subcutaneously
in an attempt to enhance the tumor-specific immunologic response. Patients will be monitored for toxicity, in
vitro and in vivo immunologic responses, and clinical anti-tumor
effects. (Protocol #9309-058)
Appendix D-66.
Drs. Peter Cassileth, Eckhard R. Podack, and Kasi Sridhar of the
University of Miami, and Niramol Savaraj of the Miami Veterans Administration
Hospital, Miami, Florida, may conduct experiments on 12 patients (≥ 18
years of age) with limited stage small cell lung cancer. Autologous tumor cells will be removed,
expanded in culture, and transduced by lipofection with the BMG-Neo-hIL2 vector
(derived from bovine papilloma virus).
The objective of this protocol is to demonstrate the safety and efficacy
of administering IL-2 transduced autologous tumor cells in an attempt to
stimulate a tumor-specific cytotoxic T lymphocyte (CTL) response, and to
determine the quantity and characteristics of the CTL that have been generated.
(Protocol #9309-053)
Appendix D-67.
Drs. Edward H. Oldfield and Zvi Ram of the National Institutes of
Health, Bethesda, Maryland, may conduct experiments on 20 patients (≥ 18
years of age) with leptomeningeal carcinomatosis. The patients will receive intraventricular or subarachnoid
injection of murine vector producing cells containing the retroviral vector,
G1Tk1SvNa. Tumor cells expressing the
herpes simplex thymidine kinase gene will be rendered sensitive to killing by
subsequent administration of ganciclovir.
Patients will be monitored for safety and anti-tumor response by
magnetic resonance imaging (MRI) and cerebral spinal fluid cytological
analysis. (Protocol #9312-059)
Appendix D-68.
Drs. Tapas K. Das Gupta and Edward P. Cohen of the University of
Illinois College of Medicine, Chicago, Illinois, may conduct experiments on 12
subjects who differ in at least 3 out of 6 alleles at the Class I
histocompatibility locus (≥ 18 years of age) with Stage IV malignant melanoma. The subjects will be immunized with a
lethally irradiated allogeneic human melanoma cell line transduced with the
human interleukin-2 expressing retroviral vector, pZipNeoSv-IL-2. Subjects will be monitored for toxicity,
induction of B and T cell antitumor responses in vitro and in vivo,
and any clinical evidence of antitumor effect. (Protocol #9309-056)
Appendix D-69A.
Dr. Michael J. Welsh of the Howard Hughes Medical Institute, Iowa City,
Iowa, may conduct experiments on 20 patients (≥ 18 years of age) with cystic
fibrosis. The investigator will
administer increasing doses of either of the two adenovirus vectors, AD2/CFTR-1
or AD2-ORF6/PGK-CFTR, to the nasal epithelium of 10 patients (1 nostril) or
maxillary sinus epithelium of 10 patients (1 maxillary sinus). Patients will be isolated for a period of 24
hours following vector administration; however, if 1 patient demonstrates
secreted virus at 24 hours, the investigator will notify the Recombinant DNA
Advisory Committee for reconsideration of the isolated period. Patients will be assessed for the safety and
efficacy of multiply administration of adenovirus vectors encoding the cystic
fibrosis transmembrane conductance regulator (CFTR) gene. (Protocol #9312-067)
Appendix D-69B.
Dr. Richard Haubrich of the University of California at San Diego
Treatment Center, San Diego, California, may conduct experiments on 25 human
immunodeficiency virus (HIV)-infected, seropositive, asymptomatic subjects (≥ 18
to ≤ 65
years of age). Subjects will receive 3
monthly intramuscular injections of the retroviral vector, N2/IIIB env/rev,
which encodes for HIV-1 IIIB env/rev proteins. The objective of the study is to induce HIV-1- specific CD8(+)
cytotoxic T lymphocyte and antibody responses in order to eliminate
HIV-infected cells and residual virus.
This Phase I/II study will evaluate acute toxicity, identify long-term
treatment effects, and evaluate the disease progression. (Protocol #9312-062)
Appendix D-70.
Dr. Mario Sznol of the National Institutes of Health, Frederick,
Maryland, may conduct experiments on 50 subjects (≥ 18 years of age) with
advanced stage melanoma. Subjects will
receive subcutaneous injections of lethally irradiated allogeneic melanoma
cells that have been transduced by lipofection with the plasmid DNA vector,
CMV-B7, derived from bovine papilloma virus to express the human B7
antigen. The B7 antigen, which binds to
the CD28 receptor of T cells, will serve as a co-stimulatory signal to elicit
an antitumor immune response. Subjects
will be monitored for induction of cytotoxic T lymphocyte, antitumor responses in
vitro and in vivo and any clinical evidence of antitumor effect.
(Protocol #9312-063)
Appendix D-71.
Dr. Joseph Rubin of the Mayo Clinic, Rochester, Minnesota, may conduct
experiments on 15 subjects with hepatic metastases from advanced colorectal
cancer (≥
18 years of age). Subjects will receive
intratumoral hepatic injections of the plasmid DNA/lipid complex, pHLA-B7/β-2 microglobulin, expressing a
heterodimeric cell surface protein consisting of the HLA-B7 histocompatibility
antigen and β-2
microglobulin. Subjects must be HLA-B7
negative. The objective of this study
is to determine a safe and effective dose of the DNA/lipid complex. Subjects will be monitored for
antigen-specific immune responses and in vivo HLA-B7 expression.
(Protocol #9312-064)
Appendix D-72.
Dr. Nicholas J. Vogelzang of the University of Chicago Medical Center,
Chicago, Illinois, may conduct experiments on 15 subjects with metastatic renal
cell carcinoma ≥ 18 years of age.
Subjects will receive intratumoral injections of the plasmid
DNA/liposome vector pHLA-B7/β-2
microglobulin, expressing a heterodimeric cell surface protein consisting of
the HLA-B7 histocompatibility antigen and β-2 microglobulin.
Subjects must be HLA-B7 negative.
Subjects will be monitored for antigen-specific immune responses and in
vivo HLA-B7 expression. (Protocol #9403-071)
Appendix D-73.
Dr. Evan M. Hersh of the Arizona Cancer Center and Drs. Emmanuel
Akporiaye, David Harris, Alison T. Stopeck, Evan C. Unger, and James A. Warneke
of the University of Arizona, Tucson, Arizona, may conduct experiments on 15
subjects with metastatic malignant melanoma ≥ 18 years of age. Subjects will receive intratumoral injections
of the plasmid DNA/liposome vector, pHLA-B7/β-2 microglobulin, expressing a heterodimeric cell surface
protein consisting of the HLA-B7 histocompatibility antigen and β-2
microglobulin. Subjects must be HLA-B7
negative. Subjects will be monitored
for antigen-specific immune responses and in vivo HLA-B7 expression.
(Protocol #9403-072)
Appendix D-74.
Dr. Ralph Freedman of MD Anderson Cancer Center, Houston, Texas, may
conduct gene marking experiments on 9 subjects with ovarian carcinoma or peritoneal
carcinomatosis (≥ 16 years of age).
Autologous CD3(+)/CD8(+) tumor infiltrating lymphocyte derived T cells
will be transduced with the retroviral vector G1Na that encodes for neoR. Subjects will receive intraperitoneal
administration of bulk expanded transduced and nontransduced T cells and
recombinant interleukin-2. Previously
documented tumor sites and normal tissues will be monitored for neoR
and the proportion of CD3(+)/CD8(+) T cells will be determined. (Protocol #9406-075)
Appendix D-75.
Drs. Helen Heslop, Malcolm Brenner, and Robert Krance of St. Jude
Children’s Research Hospital, Memphis, Tennessee, may conduct gene marking
experiments on 20 subjects undergoing autologous bone marrow transplantation
for therapy of leukemia or solid tumor (< 21 years of age). The distinguishable retroviral vectors, LNL6
and G1Na (both encoding neoR), will be used to determine the rate of
reconstitution of untreated versus cytokine expanded CD34(+) selected
autologous bone marrow cells. (Protocol
#9406-076)
Appendix D-76.
Drs. Albert Deisseroth, Gabriel Hortobagyi, Richard Champlin, and
Frankie Holmes of MD Anderson Cancer Center, Houston, Texas, may conduct
experiments on 10 fully evaluable subjects (maximum of 20 entered) with Stage
III or IV breast cancer (≥ 18 and ≤ 60 years of age).
Subjects will receive autologous CD34(+) peripheral blood cells that
have been transduced with the retroviral vector, pVMDR-1, which encodes the
multi-drug resistance gene. The
objective of this study is to evaluate the safety and feasibility of
transducing early hematopoietic progenitor cells with pVMDR-1 and to determine in
vivo selection of chemotherapy resistant hematopoietic cells. (Protocol #9406-077)
Appendix D-77.
Drs. Johnson M. Liu and Neal S. Young of the National Institutes of
Health, Bethesda, Maryland, may conduct experiments on 6 patients with Fanconi
anemia (≥
5 years of age). Subjects will receive
autologous CD34(+) cells that have been transduced with the retroviral vector,
FACC, which encodes the normal Fanconi anemia complementation group C
gene. The objective of this study is to
determine whether autologous FACC transduced hematopoietic progenitor cells can
be safely administered to subjects, the extent of engraftment, and correction
of cell phenotype. (Protocol #9406-078)
Appendix D-78.
Drs. Robert E. Sobol and Ivor Royston of the San Diego Regional Cancer
Center, San Diego, California, may conduct experiments on 15 subjects with
recurrent residual glioblastoma multi-forme (≥ 18 years of age). Subjects
will receive subcutaneous injections of autologous tumor cells that have been
lethally irradiated and transduced with the retroviral vector, G1NaCvi2.23,
which encodes for interleukin-2.
Subjects will be monitored in vitro for cellular and humoral
antitumor responses and in vivo for antitumor activity. (Protocol #9406-080)
Appendix D-79.
Dr. Alfred E. Chang of the University of Michigan Medical Center, Ann
Arbor Michigan, may conduct gene marking experiments on 15 subjects with
metastatic melanoma (≥ 18 years of age).
Subjects will undergo adoptive immunotherapy of anti-CD3/interleukin-2
activated lymph node cells that have been primed in vivo with tumor
cells that have been transduced with the retrovirus vector, GBAH4, encoding the
gene for interleukin-4. The
investigator will evaluate the antitumor efficacy and in vivo
immunological reactivity in subjects receiving adoptively transferred T cells,
and the in vitro immunological reactivities of the activated T cells
that might correlate with their in vivo antitumor function. (Protocol #9312-065)
Appendix D-80.
Dr. Robert Walker of the National Institutes of Health, Bethesda,
Maryland, may conduct gene marking experiments on 40 HIV(+) subjects (≥ 18
years of age). The investigator may
also enter an additional number of subjects (to be determined by the
investigator) who will receive a single administration of 1 x 107
HIV-specific CD8(+) cells. The
investigator will: (1) Assess the
safety and tolerance of the adoptive transfer of anti-HIV cytotoxic, syngeneic,
CD8(+) peripheral blood lymphocytes that have been transduced with the
retrovirus
vector, rkat4svgF3e-, that encodes for a universal
chimeric T cell receptor. (2) Determine
the longevity of the genetically marked CD8(+) lymphocytes in the subject's
peripheral blood. (Protocol #9403-069)
Appendix D-81.
Dr. Joseph Rosenblatt of the University of California, Los Angeles,
California, and Dr. Robert Seeger of Children’s Hospital, Los Angeles,
California, may conduct gene transfer experiments on 18 subjects with
neuroblastoma (≤ 21 years of age).
Patients at high risk of relapse with minimal or no detectable disease
following myeloablative therapy and autologous bone marrow transplantation, or
patients with progressive or persistent disease despite conventional therapy
will be serially immunized with autologous or allogeneic neuroblastoma cells
transduced to express γ
interferon. Cells will be transduced
with the retroviral vector, pHuγ-IFN,
encoding the human gene for γ
interferon and lethally irradiated prior to use as an immunogen. The objectives of the study are: (1) to
determine the maximum tolerable dose of transduced cells; (2) to determine the
local, regional, and systemic toxicities of injected cells; and (3) to
determine the antitumor response in vivo as measured by standard
clinical tests and immunocytologic evaluation of marrow metastases. (Protocol #9403-068)
Appendix D-82.
Dr. Kenneth L. Brigham of Vanderbilt University, Nashville, Tennessee,
may conduct gene transfer experiments on 10 subjects (≤ 21 years of age) in two different patient protocols (5
for each protocol). Both protocols will
use the same DNA/liposome preparations to deliver a plasmid DNA construct,
pCMV4-AAT, encoding human alpha-1 antitrypsin gene driven by a cytomegalovirus
promoter. In patients scheduled for
elective pulmonary resection, the DNA/liposome complexes will be instilled
through a fiber optic bronchoscope into a subsegment of the lung. Tissues of the lung will be obtained at the
time of surgery. Transgene expression
will be assessed by immunohistochemistry, in situ hybridization, and
Western and Northern blot analyses. The
effect of DNA/liposome complex administration on the histological appearance of
the lung will also be evaluated. In
patients with alpha-1 antitrypsin deficiency, the DNA/liposome complexes will
be instilled into the nostril.
Transgene expression will be determined in cells obtained by nasal
lavage and nasal scraping, and the time course of transgene expression will be
measured. The secretion of the alpha-1
antitrypsin protein in nasal fluid will be determined. Histological appearance of nasal mucosa will
also be examined. The study will assess
safety and feasibility of gene delivery to the human respiratory tract. (Protocol #9403-070)
Appendix D-83.
Dr. H. Kim Lyerly of Duke University Medical Center, Durham, North
Carolina, may conduct gene transfer experiments on 20 subjects with refractory
or recurrent metastatic breast cancer (≥
18 years of age). Autologous breast
cancer cells will be transduced with the DNA/liposome complex, pMP6-IL2,
containing a plasmid DNA vector derived from adeno-associated virus (AAV) that
expresses the gene for human interleukin-2.
Subjects will receive 4 subcutaneous injections of lethally irradiated
tumor cells transduced with the DNA/liposome complex prior to injection. The objective of this study is to: (1) evaluate the safety and toxicity of the
treatment, (2) determine the immunological effects, (3) determine the duration
of clinical responses, and (4) measure patient survival. (Protocol #9409-086)
Appendix D-84.
Drs. Flossie Wong-Staal, Eric Poeschla, and David Looney of the
University of California at San Diego, La Jolla, California, may conduct gene
transfer experiments on 6 subjects (≥
18 and ≤ 65 years
of age) infected with human immunodeficiency virus-1 (HIV-1). Autologous CD4(+) T lymphocytes will be
transduced ex vivo with the retroviral vector, pMJT, expressing a
hairpin ribozyme that cleaves the HIV-1 RNA in the 5' leader sequence. The transduced cells will be expanded and
reinfused into the patients. The
objectives of the study are: (1) to evaluate safety of reinfusing the
transduced lymphocytes, (2) to compare (in vivo) the kinetics and
survival of ribozyme-transduced cells with that of cells transduced with a
control vector, (3) to determine in vivo expression of the ribozyme
sequences in transduced lymphocytes, (4) to determine whether host immune
responses directed against the transduced cells will occur in vivo, and
(5) to obtain preliminary data on the effects of ribozyme gene therapy on in
vivo HIV mRNA expression, viral burden and CD4(+) lymphocyte levels. (Protocol #9309-057)
Appendix D-85.
Dr. Friedrich Schuening of the Fred Hutchinson Cancer Research Center,
Seattle, Washington, may conduct gene transfer experiments on 10 subjects (≥ 18 years of age) with Type I
Gaucher's disease. The peripheral blood
repopulating cells (mobilized by patient pretreatment with recombinant
granulocyte colony-stimulating factor) will be harvested and CD34(+) cells
selected. CD34(+) cells will be
transduced ex vivo with the retroviral vector, LgGC, that encodes human
glucocerebrosidase cDNA. Following
transduction, the transduced cells will be infused into the patient without
myeloablative treatment. The primary
endpoint of this study is to examine the safety of infusing CD34(+) cells
transduced with the human glucocerebrosidase cDNA. Patients will be monitored for persistence and expression of the
glucocerebrosidase gene in hematopoietic cells. (Protocol #9312-061)
Appendix D-86.
Dr. Terence R. Flotte of the Johns Hopkins Children's Center, Baltimore,
Maryland, may conduct gene transfer experiments on 16 subjects (≥ 18 years of age) with mild
cystic fibrosis (CF). An
adeno-associated virus (AAV) derived vector, encoding cystic fibrosis
transmembrane conductance regulator (CFTR) gene, (tgAAVCF), will be
administered to nasal (direct) and airway (bronchoscope) epithelial cells. This is a dose escalation study involving 8
cohorts. Each subject will receive both
intranasal and bronchial administration of the adenoviral vector at 4
escalating doses. Nasal doses will
range between 1 x 106 and 1 x 109 pfu. Lung administration will range between 1 x
107 and 1 x 1010 pfu.
The primary goal of the study is to assess the safety of vector
administration. Respiratory and nasal
epithelial cells will be evaluated for gene transfer, expression, and
physiologic correction. (Protocol
#9409-083)
Appendix D-87.
Drs. Jeffrey M. Isner and Kenneth Walsh of St. Elizabeth's Medical
Center, Tufts University, Boston, Massachusetts, may conduct gene transfer
experiments on 12 subjects (≥ 40
years of age) with peripheral artery disease (PAD). A plasmid DNA vector, phVEGF165, encoding the human gene for
vascular endothelial growth factor (VEGF) will be used to express VEGF to
induce collateral neovascularization. Percutaneous
arterial gene transfer will be achieved using an angioplasty catheter with a
hydrogel coated balloon to deliver the plasmid DNA vector to the artery. The objectives of the study are: (1) to determine the efficacy of arterial
gene therapy to relieve rest pain and/or heal ischemic ulcers of the lower extremities
in patients with PAD; and (2) to document the safety of the phVEGF arterial
gene therapy for therapeutic angiogenesis.
Subjects will undergo anatomic and physiologic examination to determine
the extent of collateral artery development following phVEGF arterial gene
therapy. (Protocol #9409-088)
Appendix D-88A.
Dr. Ronald G. Crystal of New York Hospital-Cornell Medical Center, New
York, New York, may conduct gene transfer experiments on 26 patients (≥ 15 years of age) with cystic
fibrosis (CF). A replication deficient
recombinant adenovirus vector will be used to transduce epithelial cells of the
large bronchi with the E1/E3 deleted type 5 adenovirus vector, AdGVCFTR.10,
which encodes the human cystic fibrosis transmembrane conductance regulator
(CFTR) gene. The objective of this
study is to define the safety and pharmacodynamics of CFTR gene expression in
airway epithelial cells following single administration of escalating doses to
the vector. If single administration is
determined to be safe, subjects will undergo repeat administration to localized
areas of the bronchi. (Protocol
#9409-085)
Appendix D-88B.
Drs. Eric J. Sorscher and James L. Logan of the University of Alabama,
Birmingham, Alabama, may conduct gene transfer experiments on 9 subjects (≥18 years of age) with cystic
fibrosis (CF). The normal human cystic
fibrosis transmembrane conductance regulator (CFTR) gene will be expressed by a
plasmid DNA vector, pKCTR, driven by the simian virus-40 (SV40) early gene
promoter. The CFTR DNA construct will
be delivered by cationic liposome-based gene transfer to nasal epithelial
cells. The objectives of the study are
to: (1) evaluate the safety of
lipid-mediated gene transfer to nasal epithelial cells (including local
inflammation and mucosal tissue); and (2) evaluate efficacy as determined by
correction of the chloride ion transport defect, and wild-type CFTR mRNA and
protein expression. (Protocol #9312-066)
Appendix D-89.
Dr. Steven M. Albelda of the University of Pennsylvania Medical Center,
Philadelphia, Pennsylvania, may conduct gene transfer experiments on 12
subjects with advanced mesothelioma.
The adenovirus vector encoding the Herpes simplex virus thymidine
kinase (HSV-TK) gene, H5.020RSVTK, will be administered through a chest tube to
the pleural cavity. Tumor biopsies will
be assayed for gene transfer and expression.
Subjects will be monitored for immunological responses to the adenovirus
vector. Ganciclovir will be
administered intravenously 14 days following vector administration. The primary objective of this Phase I study
is to evaluate the safety of direct adenovirus vector gene delivery to the
pleural cavity of patients with malignant melanoma. (Protocol #9409-090)
Appendix D-90.
Drs. Jeffrey Holt and Carlos B. Arteaga of the Vanderbilt University,
Nashville, Tennessee, may conduct gene transfer experiments on 10 female
patients (over 18 years of age) with metastatic breast cancer. Patient
effusions from pleura or peritoneum will be drained and the fluid will be replaced
with supernatant containing the retroviral vectors, XM6:antimyc or XM6:antifos,
which express c-myc and c-fos antisense sequences, respectively,
under the control of a mouse mammary tumor virus promoter. The objectives of this study are to: (1)
assess uptake and expression of the vector sequences in breast cancer
cells present in pleural and peritoneal fluids, and determine if this
expression is tumor specific, (2) assess the safety of localized administration
of antisense retroviruses, and (3) monitor subjects for clinical evidence of
antitumor response. (Protocol #9409-084)
Appendix D-91.
Dr. Jack A. Roth of MD Anderson Cancer Center, Houston, Texas, may
conduct gene transfer experiments on 14 non-small cell lung cancer subjects (≥ 18 and ≤ 80 years of age) who have
failed conventional therapy and who have bronchial obstruction. LNSX-based retroviral vectors containing the
β-actin promoter will be
used to express: (1) the antisense RNA of the K-ras oncogene (LN-K-rasB),
and (2) the wildtype p53 tumor suppressor gene (LNp53B). Tumor biopsies will be obtained to
characterized K-ras and p53 mutations. Relative to their specific mutation, subjects will undergo
partial endoscopic resection of the tumor bed followed by bronchoscopic
administration of the appropriate retrovirus construct. The objective of this study is to evaluate
the safety and efficacy of intralesional administration of LN-K-rasB and
LNp53 retrovirus constructs. (Protocol #9403-031)
Appendix D-92.
Drs. Robert E. Sobol and Ivor Royston of the San Diego Regional Cancer
Center, San Diego, California, may conduct gene transfer experiments on 12
subjects (≥ 18 years of
age) with metastatic colon carcinoma.
The autologous skin fibroblasts will be transduced with the retroviral
vector, LNCX/IL-2, which encodes the gene for human interleukin-2 (IL-2). In this dose-escalation study, subjects will
receive subcutaneous injections of lethally irradiated autologous tumor
cells. The objectives of the study are
to: (1) evaluate the safety of subcutaneous administration of LNCX/IL-2
transduced fibroblasts, (2) determine in vivo antitumor activity, and
(3) monitor cellular and humoral antitumor responses. (Protocol #9312-060)
Appendix D-93.
Dr. Michael Lotze of the University of Pittsburgh, Pittsburgh,
Pennsylvania, may conduct gene transfer experiments on 18 subjects (≥ 18 years of age) with
advanced melanoma, 6 with T-cell lymphoma, breast cancer, or head and neck
cancer. Subjects should have accessible
cutaneous tumors, and have failed standard therapy. Over 4 weeks, subjects will receive a total of 4 intratumoral
injections of autologous fibroblasts transduced with the retrovirus vector,
TFG-hIL-12-Neo. This vector, which
consists of the murine MFG backbone, expresses both the p35 and p40 subunits of
interleukin-12 (IL-12) and the neoR selection marker. The objectives of the study are to: (1) define the local and systemic toxicity
associated with peritumoral injections of gene-modified fibroblasts, (2) examine
the local and systemic immunomodulatory effects of these injections, and (3)
evaluate clinical antitumor efficacy.
(Protocol #9406-081)
Appendix D-94.
Drs. Evan Hersh, Emmanuel Akporiaye, David Harris, Alison Stopeck, Evan
Unger, James Warneke, of the Arizona Cancer Center, Tucson, Arizona, may
conduct gene transfer experiments on 25 subjects (≥ 18 years of age) with solid malignant tumors or
lymphomas. A plasmid DNA/lipid complex
designated as VCL-1102 (IL-2 Plasmid DNA/DMRIE/DOPE) will be used to transduce
the human gene for interleukin-2 (IL-2).
Patients with advanced cancer who have failed conventional therapy will
undergo a procedure in which VCL-1102 is injected directly into the tumor mass
to induce tumor-specific immunity. The
objectives of the study are to: (1)
determine safety and toxicity associated with escalating doses of VCL-1102; (2)
confirm IL-2 expression in target cells; (3) determine biological activity and
pharmacokinetics; and (4) determine whether IL-2 expression stimulates tumor
regression in subjects with metastatic malignancies. (Protocol #9412-095)
Appendix D-95.
Drs. Richard Morgan and Robert Walker of the National Institutes of
Health, Bethesda, Maryland, may conduct gene transfer experiments on 48 human
immunodeficiency virus (HIV) seropositive subjects (≥ 18 years of age).
This Phase I/II study involves identical twins (one HIV seropositive and
the other HIV seronegative). CD4(+) T
cells will be enriched following apheresis of the HIV seronegative twin,
induced to polyclonal proliferation with anti-CD3 and recombinant IL-2,
transduced with either the LNL6/NeoR or G1Na/NeoR, and
transduced with up to 2 additional retroviral vectors (G1RevTdSN and/or
GCRTdSN(TAR)) containing potentially therapeutic genes (antisense TAR and/or
transdominant Rev). These T cell
populations will be expanded 10 to 1,000 fold in culture for 1 to 2 weeks and
reinfused into the HIV seropositive twin.
Subjects will receive up to 4 cycles of treatment using identical or
different combinations of control and anti-HIV retrovirus vectors. The relative survival of these transduced T
cell populations will be monitored by vector-specific polymerase chain
reaction, while the subjects' functional immune status is monitored by standard
in vitro and in vivo assays.
(Protocol #9503-103)
Appendix D-96.
Dr. Harry L. Malech of the National Institutes of Health, Bethesda,
Maryland, may conduct gene transfer experiments on 2 subjects ≥ 18 years of age (with or
without concurrent serious infection), and 3 subjects ≥ 18 years of age (with or without concurrent serious
infection) or minors 13-17 years of age who have concurrent serious infection
who have chronic granulomatous disease (CGD).
CGD is an inherited immune deficiency disorder in which blood
neutrophils and monocytes fail to produce antimicrobial oxidants (p47phox
mutation) resulting in recurrent life-threatening infections. Subjects will undergo CD34(+) mobilization
with granulocyte colony stimulating factor (G-CSF). These CD34(+) cells will be transduced with the retrovirus
vector, MFG-S-p47phox, which encodes the gene for normal p47phox. The objectives of this study are to: (1) determine the safety of administering
MFG-S-p47phox transduced CD34(+) cells, and (2) demonstrate
increased functional oxidase activity in circulating neutrophils. (Protocol #9503-104)
Appendix D-97.
Drs. Chris Evans and Paul Robbins of the University of Pittsburgh,
Pittsburgh, Pennsylvania, may conduct gene transfer experiments on 6 subjects (≥ 18 and ≤ 76 years of age) with rheumatoid
arthritis. Rheumatoid arthritis is a
chronic, progressive disease thought to be of autoimmune origin. A gene encoding an interleukin-1 receptor
antagonist protein (IRAP) will be delivered to the rheumatoid
metacarpal-phalangeal joints to determine the autoimmune reactions can be
interrupted. The vector construct,
DFG-IRAP, is based on the MFG murine retrovirus vector backbone, and encodes
the human IRAP gene. Synovial fibroblasts
will be generated from the rheumatoid arthritic joint tissue obtained from
patients who are scheduled to undergo surgery.
The fibroblasts will be transduced with the DFG-IRAP vector, and the
transduced cells injected into the synovial space. The synovial fluid and joint material will be collected 7 days
later to determine the presence and location of the transduced synovial
fibroblasts and the level of IRAP in the joint fluid. (Protocol 9406-074)
Appendix D-98.
Dr. R. Scott McIvor of the University of Minnesota, Minneapolis,
Minnesota, may conduct gene transfer experiments on 2 children with purine
nucleoside phosphorylase (PNP) deficiency.
PNP deficiency results in severe T-cell immunodeficiency, an autosomal
recessive inherited disease which is usually fatal in the first decade of
life. Autologous peripheral blood lymphocytes
will be cultured in an artificial capillary cartridge in the presence of
anti-CD3 monoclonal antibody and interleukin-2 and transduced with the
retroviral vector, LPNSN-2, encoding human PNP. Subjects will undergo bimonthly intravenous administration of
transduced T cells for a maximum of 1 year.
The objectives of the study are to determine: (1) the safety of intravenous administration of transduced T
cells in children with PNP deficiency, (2) the efficiency of PNP gene transfer
and duration of gene expression in vivo, and (3) the effect of PNP gene
transfer on immune function. (Protocol
#9506-110)
Appendix D-99.
Drs. Nikhil C. Munshi and Bart Barlogie of the University of Arkansas
School for Medical Sciences, Little Rock, Arkansas, may conduct gene transfer
experiments on 21 subjects (>18 and <65 years of age) with relapsed or
persistent multiple myeloma who are undergoing T cell depleted allogeneic bone
marrow transplantation. Donor
peripheral blood lymphocytes will be cultured in vitro with interleukin-2 and
anti-CD3 monoclonal antibody. T cell
depleted lymphocytes will be transduced with the retroviral construct,
G1Tk1SvNa.7, which encodes the Herpes simplex virus thymidine kinase (HSV-TK)
gene. The transduced cells will be
reinfused. In this dose escalation
study, 3 subjects will undergo cell-mediated gene transfer per cohort (maximum
of 5 cohorts) until Grade III or IV Graft versus Host Disease (GVHD) is
observed. A maximum of 6 additional patients
may be entered at that maximum tolerated dose.
The objectives of this study are to determine the: (1) safety of transduced donor cell
infusions, (2) effectiveness of donor cell infusions in decreasing the effects
of severe GVHD, (3) effectiveness of donor cell infusions in prolonging multiple
myeloma remission, and (4) effectiveness of ganciclovir in eliminating donor
cells for the purpose of preventing the depletion of erythrocytes. (Protocol #9506-107)
Appendix D-100.
Dr. Wayne A. Marasco of Dana-Farber Cancer Institute, Boston, Massachusetts,
may conduct gene transfer experiments on 6 subjects (≥ 18 and ≤ 65
years of age) with human immunodeficiency virus type-1 (HIV-1). Autologous lymphocytes from asymptomatic
subjects will be transduced ex vivo with a retroviral vector, LNCs105,
encoding the sFv105 antibody specific for the HIV-1 envelope protein. An identical aliquot will be simultaneously
transduced with a control retroviral vector lacking the sFv105 cassette. Transduced cells will be reinfused into
patients and the differential survival of both populations of CD4+ lymphocytes
compared. The objective of the study is
to determine whether the intracellular expression of a human single chain
antibody against HIV-1 envelope glycoprotein gp160 that blocks gp160 processing
and the production of infectious virions can safely prolong the survival of
CD4(+) lymphocytes in HIV-1-infected subjects.
(Protocol #9506-111)
Appendix D-101.
Dr. Henry Dorkin of the New England Medical Center, Boston,
Massachusetts, and Dr. Allen Lapey of Massachusetts General Hospital, Harvard
Medical School, Boston, Massachusetts, propose to conduct gene transfer
experiments on 16 subjects (≥ 18
years of age). An E1/partial
E4-deleted, replication-deficient, type 2 adenovirus vector, AD2/CFTR-2, will
be used to deliver the human cystic fibrosis transmembrane conductance
regulator (CFTR) gene by aerosol administration (nebulization) to the lung of
CF patients. Aerosol administration
will be initiated only after initial safety data has been obtained from the
lobar administration protocol (#9409-091).
This is a single administration dose-escalation study in which subjects
will receive between 8 x 106 and 2.5 x 1010 pfu. Subjects will be assessed for evidence of
adverse, systemic, immune, inflammatory, or respiratory effects in response to
AD2/CFTR-2. Subjects will be monitored
for virus shedding and transgene expression.
Health care workers present in the facility will be required to sign an
Informed Consent document regarding the possibility of virus transmission. (Protocol #9412-074)
Appendix D-102.
Drs. Charles J. Link and Donald Moorman of the Human Gene Therapy
Research Institute, Des Moines, Iowa, may conduct gene transfer experiments on
24 female subjects (≥ 18
years of age) with refractory or recurrent ovarian cancer. Subjects will undergo intraperitoneal
delivery (via Tenkhoff catheter) of the vector producing cells (VPC),
PA317/LTKOSN.2. These VPC express the Herpes
simplex virus thymidine kinase (HSV-TK) gene which confers sensitivity to
killing by the antiviral drug, ganciclovir (GCV). The LTKOSN.2 retrovirus vector is based on the LXSN
backbone. Two weeks following
intraperitoneal delivery of the VPC, subjects will receive 5 mg/kg intravenous
GCV twice daily for 14 days. Subjects
will receive between 1 x 105 and 1 x 108 VPC/kg in this
dose escalation study. Subjects will be
evaluated by X-ray and peritoneoscopy of the abdomen for evidence of clinical
response. The objectives of this study
are to determine the safety of intraperitoneal VPC administration. (Protocol #9503-100)
Appendix D-103.
Dr. David T. Curiel of the University of Alabama, Birmingham, Alabama,
may conduct gene transfer experiment of 15 subjects (≥ 18 years of age) with metastatic colorectal cancer. Subjects will receive intramuscular
injection of the polynucleotide vaccine, pGT63, which is a plasmid DNA vector
expressing carcinoembryonic antigen (CEA) and hepatitis B surface antigen
(HBsAg). The objectives of the study
are to: (1) characterize the immune
response to CEA and HBsAg following a single intramuscular injection and
following 3 consecutive intramuscular injections, and (2) determine the safety
of intramuscular injection of the plasmid DNA vector at doses ranging between
0.1 to 1.0 milligrams (single dose) and 0.9 to 3.0 milligrams (total
multidose). (Protocol #9506-073)
Appendix D-104.
Dr. Chester B. Whitley of the University of Minnesota, Minneapolis,
Minnesota, may conduct gene transfer experiments on two adult subjects (18
years of age or older) with mild Hunter syndrome (Mucopolysaccharidosis Type
II). The autologous peripheral blood
lymphocytes will be transduced ex vivo with the retroviral vector, L2SN,
encoding the human cDNA for iduronate-2-sulfatase (IDS). The transduced lymphocytes will be reinfused
into the patients on a monthly basis.
The study will determine the frequency of peripheral blood lymphocyte
transduction and the half-life of the infused cells. Evaluation of patients will include measurement of blood levels
of IDS enzyme, assessment of metabolic correction by urinary glycosaminoglycan
levels, clinical response of the disease, and monitoring for potential
toxicity. This Phase I study is to
demonstrate the safety of the L2SN-mediated gene therapy and to provide a
preliminary evaluation of clinical efficacy.
(Protocol #9409-087)
Appendix D-105.
Drs. James Economou, John Glaspy, and William McBride of the University
of California, Los Angeles, California, may conduct gene transfer experiments
on 25 subjects (≥ 18
years of age) with metastatic melanoma.
The protocol is an open label, Phase I trial to evaluate the safety and
immunological effects of administering lethally irradiated allogeneic and
autologous melanoma cells transduced with the retroviral vector, IL-7/HyTK,
which encodes the gene for human interleukin-7 (IL-7). Subjects will receive 1 x 107
irradiated unmodified autologous tumor cells in combination with escalating
doses of IL-7/HyTK transduced allogeneic melanoma cells (M24 cell line). The number of M24 cells administered will be
adjusted based on the level of IL-7 expression. Subjects will receive 3 biweekly subcutaneous injections of M24
cells expressing 10, 100, or 1000 nanograms of IL-7/hour in vivo. A final cohort of 5 subjects will receive
IL-7/HyTK transduced autologous cells.
Subjects will be monitored for antitumor activity by skin tests, biopsy
analysis, tumor-specific antibody activity, and cytotoxic T lymphocyte
precursor evaluation. Non-immunologic
parameters will also be monitored. (Protocol #9503-101)
Appendix D-106.
Dr. Jack A. Roth, MD Anderson Cancer Center, may conduct gene transfer
experiments on 42 subjects (≥ 18
years of age) with refractory non-small cell lung cancer (NSCLC). Subjects will receive direct intratumoral injection
of a replication-defective type 5 adenovirus vector, AD5CMV-p53, to deliver the
normal human p53 tumor suppressor gene.
The E1 region of AD5CMV-p53 has been replaced with a p53
expression cassette containing the human cytomegalovirus promoter (CMV). Subjects will be divided into 2 treatment
groups: (1) 21 subjects will receive
Ad5CMV-p53 alone, and (2) 21 subjects will receive Ad5CMV-p53 in combination
with cisplatin. Following vector
administration, subjects will be isolated for 96 hours during which time,
assays will be conducted to demonstrate the lack of shedding of adenovirus
vector. The objectives of this study
are determine: (1) the maximum
tolerated dose of AD5CMV-p53, (2) qualitative and quantitative toxicity related
to vector administration, and (3) biologic activity.
Prior to administration, adenovirus vector stocks will be
screened for p53 mutants using the SAOS osteosarcoma cell assay that was
submitted by Dr. Roth on June 23, 1995.
This biologic assay compares the activity of a standard stock of Adp53
vector to the activity of newly produced stocks. The standard stock of Adp53 will be defined as mediating cell
death in 100% of SAOS cells (human osteosarcoma cell line with homozygous p53 deletion)
at an MOI of 50:1 (titer > 5
x 1010) on day 5 of culture.
The sensitivity of the assay for detecting inactive (presumed mutant)
Adp53 vector will be determined by adding increasing amounts of Adluc (control
adenovirus vector containing the luciferase gene) to the Adp53 stock to determine
the percentage of inactive vector required to decrease growth inhibition of
SAOS cells mediated by Adp53. The test
lot of Adp53 will be tested for its ability to inhibit SAOS in a 5 day assay. Significant loss of inhibitory activity
compared with the standard would indicate an unacceptable level of inactive
(presumed mutant) vector. (Protocol #9406-079)
Appendix D-107A.
Dr. Gary Clayman. M.D. Anderson Cancer Center, Houston, Texas, may
conduct gene transfer experiments on 21 subjects (≥ 18 years of age) with refractory squamous cell carcinoma
of the head and neck. Subjects will
receive direct intratumoral injection of a replication-defective type 5
adenovirus vector, AD5CMV-p53, to deliver the normal human p53 tumor
suppressor gene. The E1 region of
AD5CMV-p53 has been replaced with a p53 expression cassette containing
the human cytomegalovirus promoter (CMV).
Subjects will be divided into 2 treatment groups: (1) those with
non-resectable tumors, and (2) those with surgically accessible tumors. Subjects will receive multiple injections of
vector in each dose-escalation cohort.
Following vector administration, subjects will be isolated for 48 hours
during which time, assays will be conducted to demonstrate the lack of shedding
of adenovirus vector. The objectives of
the study are to determine: (1) the maximum tolerated dose of AD5CMV-p53, (2)
qualitative and quantitative toxicity related to vector administration, and (3)
biologic activity.
Prior to administration, adenovirus vector stocks will be
screened for p53 mutants using the SAOS osteosarcoma cell assay that was
submitted by Dr. Roth on June 23, 1995.
This biologic assay compares the activity of a standard stock of Adp53
vector to the activity of newly produced stocks. The standard stock of Adp53 will be defined as mediating cell
death in 100% of SAOS cells (human osteosarcoma cell line with homozygous p53
deletion) at an MOI of 50:1 (titer >
5x1010) on day 5 of culture.
The sensitivity of the assay for detecting inactive (presumed mutant)
Adp53 vector will be determined by adding increasing amounts of Adluc to the
Adp53 stock to determine the percentage of inactive vector required to decrease
growth inhibition of SAOS cells mediated by Adp53. The test lot of Adp53 will be tested for its ability to inhibit
SAOS in a 5 day assay. Significant loss
of inhibitory activity compared with the standard would indicate an
unacceptable level of inactive (presumed mutant) vector. (Protocol #9412-096)
Appendix D-107B.
Drs. Bernard A. Fox and Walter J. Urba of Earle A. Chiles Research
Institute, Providence Medical Center, Portland, Oregon, may conduct gene
transfer experiments on 18 subjects (≥
18 years of age) with metastatic renal cell carcinoma or melanoma. Autologous tumor cells will be surgically
removed, transduced in vitro with the cationic liposome plasmid vector,
VCL-1005, which encodes human leukocyte antigen (HLA)-B7 and beta-2
microglobulin. Subjects will receive
subcutaneous injection of lethally irradiated transduced cells in one limb. The contralateral limb will be injected with
lethally irradiated untransduced tumor cells in combination with Bacille
Calmette-Guerin (BCG). Approximately 21
days following tumor cell injection, subjects will undergo lymphadenectomy for
subsequent in vitro expansion of anti-CD3 activated lymphocytes. Activated lymphocytes will be adoptively
transferred on approximately day 35 in combination with a 5-day course of
interleukin-2 (IL-2). On approximately
day 45, subjects will receive a second cycle of IL-2. The objectives of this study are to determine: (1) the safety of administering anti-CD3
activated antitumor effector T cells in draining lymph nodes, and (2) whether
HLA-B7/β-2 gene transfer
augments the sensitization of anti-tumor effector T-cells in draining lymph
nodes. (Protocol 9506-108)
Appendix D-108.
Dr. Mitchell S. Steiner, University of Tennessee, Memphis, Tennessee,
and Dr. Jeffrey T. Holt, Vanderbilt University School of Medicine, Nashville,
Tennessee, may conduct gene transfer experiments on 15 male subjects (35 to 75
years of age) with metastatic prostate cancer.
Malignant cells obtained from advanced prostate cancer subjects have
been demonstrated to express high levels of the protooncogene c-myc in vivo. The mouse mammary tumor virus (MMTV) long
terminal repeat (LTR) is expressed at high levels in prostate tissue. Following removal of malignant cells via
biopsy, subjects will receive a single transrectal ultrasound-guided
intraprostate quadrant injection of the retrovirus vector, XM6:MMTV-antisense
c-myc, for 4 consecutive days at the site of the original biopsy. The objectives of this Phase I study are
to: (1) quantitatively assess the
uptake and expression of XM6:MMTV-antisense c-myc by prostate cancer cells in
vivo, (2) determine whether c-myc gene expression is prostate
tumor-specific, (3) assess safety of intraprostate injection of
XM6:MMTV-antisense c-myc, and (4) biologic efficacy (antisense inhibition of
tumor growth). (Protocol #9509-123)
Appendix D-109.
Drs. Ronald G. Crystal, Edward Hershowitz, and Michael Lieberman, New
York Hospital-Cornell Medical Center, New York, New York, may conduct gene
transfer experiments on 18 subjects (18 to 70 years of age) with metastatic
colon carcinoma with liver metastases.
In this Phase I dose-escalation study, subjects will receive computed
tomography (CT)-guided intratumoral injections of the adenovirus vector, AdGVCD.10,
into the same hepatic metastasis in 4 equal volumes (100 microliters), each
with a separate entry into the liver.
This dosage schedule will be performed on Days 1 and 7. 5-fluorocytosine (200 milligrams/
kilogram/24 hours) will be administered orally in 4 equal doses starting on day
2 and continuing through the time of laparotomy. The objectives of this study are to: (1) determine the dose-dependent toxicity of direct
administration of AdGVCD.10 to hepatic metastases
combined with oral administration of 5-fluorocytosine, (2) quantitatively
assess transfer and expression of the cytosine deaminase gene in target cells,
and (3) determine the biologic effects of direct ADGVCD.10
administration to hepatic metastases. (Protocol #9509-125)
Appendix D-110.
Drs. Andres Berchuck and H. Kim Lyerly of Duke University Medical Center,
Durham, North Carolina, may conduct gene transfer experiments on 18 subjects (≥ 18 years of age) with
refractory metastatic ovarian cancer.
Autologous tumor cells obtained from ascites or surgically removed tumor
will be transduced with the cationic liposome vector, PMP6A-IL2, that contains
an adeno-associated virus derived plasmid DNA, a cytomegalovirus (CMV)
promoter, and interleukin-2 (IL-2) complementary DNA (cDNA). In this dose-escalation study, subjects will
undergo 4 cycles of intradermal injections (thigh or abdomen) of ex vivo
transduced, lethally irradiated tumor cells in an attempt to induce an
antitumor response. The objectives of
the study are to evaluate: (1) the
safety of intradermally injected transduced cells, and (2) antitumor response
following therapy. (Protocol #9506-110)
Appendix D-111.
Drs. Stephen L. Eck and Jane B. Alavi of the University of Pennsylvania
Medical Center, Philadelphia, Pennsylvania, may conduct gene transfer
experiments on 18 subjects (>18 years of age) with malignant glioma. The adenovirus vector encoding the Herpes
simplex virus thymidine kinase (HSV-TK) gene, H5.020RSVTK, will be injected
by a stereotactic guided technique into brain tumors. Afterwards, the patients will receive systemic ganciclovir (GCV)
treatment. Patients eligible to undergo
a palliative debulking procedure will receive the same treatment followed by
resection on day 7, and a second dose of the vector intra-operatively. Brain tissues removed by resection will be
analyzed for adenovirus infection, transgene expression, and signs of
inflammation. The size and metabolic
activity of tumors will be monitored by scanning with magnetic resonance
imaging and positron emission tomography.
The objective of the study is to evaluate the overall safety of this
treatment and to gain insight into the parameters that may limit the general
applicability of this approach.
(Protocol #9409-089)
Appendix D-112.
Drs. Robert Grossman and Savio Woo of the Baylor College of Medicine
& Methodist Hospital, Houston, Texas, may conduct gene transfer experiments
on 20 subjects (≥ 18
years of age) with refractive central nervous system malignancies. Subjects will receive stereotaxic injections
of a replication-defective, type 5 E1/E3-deleted adenovirus vector, ADV/RSV-tk,
to deliver the Herpes simplex virus thymidine kinase (HSV-TK) gene to
tumor cells. Expression of the HSV-TK
gene is driven by a Rous sarcoma virus long terminal repeat (RSV-LTR). Subjects will receive a single time-course
of intravenous ganciclovir (GCV) (14 consecutive days) following vector
administration. Following demonstration
of safety with the initial starting dose of 1 x 108 particles in 5
subjects, additional cohorts will receive between 5 x 108 and 1.5 x
109 particles. Each cohort
will be monitored for toxicity for one month before administration of the next
higher dose to subsequent cohorts.
Subjects will be monitored for evidence of clinical efficacy by magnetic
resonance imaging and/or computer tomography scans. The primary objective of this Phase I study is to determine the
safety of vector administration.
(Protocol #9412-098)
Appendix D-113.
Drs. Gabriel N. Hortobagyi, Gabriel Lopez-Berestein, and Mien-Chie Hung,
of the University of Texas MD Anderson Cancer Center, Houston, Texas, may
conduct gene transfer experiments on a maximum of 24 adult patients (12 for
each cancer) with metastatic breast or ovarian carcinoma. Overexpression of the HER-2/neu
oncogene occurs in 30% of ovarian and breast cancers, and it is associated with
enhanced metastatic potential, drug resistance, and poor survival. The E1A gene of the adenovirus type 5
functions as a tumor suppressor gene when transfected into cancer cells which
overexpress the HER-2/neu oncogene.
E1A expression induces down regulation of the level of the HER-2/neu
oncoprotein by a transcriptional control mechanism. A plasmid, pE1A, encoding the adenovirus E1A gene with its own
promoter will be administered as a DNA/lipid complex via the intraperitoneal or
intrapleural route. The objectives of
the study are: (1) to determine E1A
gene transduction into malignant cells after the administration of E1A/lipid
complex by intrapleural or intraperitoneal administration, (2) to determine
whether E1A gene therapy can down-regulate HER-2/neu expression after
intrapleural or intraperitoneal administration, (3) to determine the maximum
biologically active dose (MBAD), or the maximum tolerated dose (MTD) of
E1A/lipid complex, (4) to determine the toxicity and tolerance of E1A/lipid complex
administered into the pleural or peritoneal space, and to assess the
reversibility of such toxicity, and (5) to evaluate tumor response. (Protocol #9512-137)
Appendix D-114.
Drs. Keith L. Black and Habib Fakhrai of the University of California, Los
Angeles, California, may conduct gene transfer experiments on 12 subjects (≥ 18 years of age) with
glioblastoma multiform. An Epstein-Barr
virus (EBV) based plasmid vector, pCEP-4/TGF- β2 antisense, encoding antisense RNA will be used to
inhibit TGF- β2
production. Tumor samples obtained from
the patients at the time of clinically indicated surgery will be grown in
culture to establish a cell line for each patient. The patients' tumor cells will be genetically altered with the
pCEP-4/TGF-β2 vector to
inhibit their secretion of TGF-β. Following completion of the traditional post
surgical radiation therapy, the first cohort of patients will receive, at 3
week intervals, 4 injections of 5 x 106 irradiated gene modified
autologous tumor cells. Subsequently,
in dose escalation studies, the second cohort will receive 1 x 107
cells, and the third cohort, 2 x 107 cells. The results of this Phase I trial will be
used to assess the safety of this form of gene therapy and may provide
preliminary data to evaluate the potential utility of TGF- β2 antisense gene therapy in
the management of gliomas. (Protocol
#9512-138)
Appendix D-115.
Dr. Ronald G. Crystal of New York Hospital-Cornell Medical Center, New
York, New York, may conduct gene transfer experiments on a total of 21 (with an
option for an additional 5) normal males and female subjects, age ≥ 18 years. Replication-deficient adenovirus (Ad) vector
previously has been used in a number of human gene therapy strategies to
transfer genes in vivo for therapeutic purposes. The purpose of this protocol is to
characterize the local (skin), systemic (blood), and distant compartment (lung)
immunity in normal individuals after intradermal administration of a
replication deficient Ad5-based vector, named AdGVCD.10, carrying
the gene coding for the E. coli enzyme, cytosine deaminase (CD). Following intradermal administration of the
vector to normal individuals, the skin, blood, and lung immune responses to the
Ad vector and CD transgene will be evaluated over time. This vector has been safety administered
intrahepatically ten times to five individuals with colon carcinoma. No adverse effects in Protocol #9509-125
have been observed. The present
protocol will yield insights into normal human immune responses to both the Ad
vector, as well as to a heterologous (i.e., non-human) gene product (CD). Note: This study is designed to answer basic
biological questions regarding characterization of the immune responses to such
vectors that have been previously documented.
(Protocol #9701-171)