Last Updated: 09/13/10
Dana-Farber/Harvard Cancer Center
Overall Abstract
Principal Investigator:
Kenneth C. Anderson, M.D.
The Dana Farber/Harvard Cancer Center (DF/HCC) multiple myeloma (MM) SPORE consists of 5 Research Projects and 4 Cores, as well as the Career Development and Developmental Research Programs. It capitalizes on the complementary strengths of the research, clinical expertise and facilities of the Harvard affiliated institutions including Dana Farber Cancer Institute, Harvard Medical School, Harvard School of Public Health, as well as the Mayo Clinic, the Cornell University Weil Medical College; and the University of Arizona Medical School. This SPORE represents a collaborative effort between MM research groups from these institutions with a long-standing commitment and track record of basic and clinical scientific interactions and cooperation. We have established a collaborative effort, both in preclinical cellular and molecular studies and in joint clinical protocols. The group as a whole has a long-term commitment to translational MM research, with the necessary administrative, basic science, and clinical infrastructure. At these well established centers, more than 750 new patients with MM are evaluated annually, as well as 10,000 outpatient visits for established patients with plasma cell dyscrasias. The spectrum of diseases evaluated spans from monoclonal gammopathy of unclear significance to plasma cell leukemia. Each center has appropriate scientific and institutional review boards, as well as protocol audit and quality control centers, to conduct cutting edge translational research. There are presently more than 50 active protocols evaluating therapies including novel drugs, immune treatments, improved stem cell transplantation, and supportive therapies in MM. This large combined patient base assures rapid accrual and evaluation of the therapeutic efficacy of novel agents identified in this program. Success of both the preclinical and clinical components of this Program will be dependent upon synergy and communication between these centers. To assure this end, we have set up an Internet site that allows access to all the Principal Investigators to the preclinical data generated in joint research efforts. Similarly, data from the joint clinical protocol trials will also be deposited in this secure web site to allow a seamless and uniform conduct of clinical studies at these sites. Currently there is systematic quality-controlled exchange of bone marrow and blood samples for correlative basic laboratory studies. The overall theme of the DF/HCC myeloma SPORE is to identify and evaluate novel targeted therapies. The translational nature of the SPORE is highlighted by the fact that most of our projects have emanated from clinical studies from the outset Specific Projects are (1) Proteasome-Directed Novel Myeloma Therapies; (2) Targeting Telomere Expansion Mechanisms For Myeloma Therapy; (3) MUC1 as a Therapeutic Target in Multiple Myeloma; (4) Novel Therapeutics Targeting Genetic Abnormalities in MM; and (5) Molecular Markers of Evolution from MGUS To Myeloma. Core resources include Administrative and Communication Core (1), Tissue Core (2), and Biostatistics Core (3). This Program therefore represents the integrated efforts of institutions with a unique and long track record of basic and clinical research expertise in MM, now joining together to more rapidly move rational novel targeted therapies from the laboratory to clinical protocols to improve patient outcome in MM.
Project 1: Proteasome-Directed Novel Myeloma Therapies
Principal Investigator:
Kenneth C. Anderson — DFCI
Co-Principal Investigator:
Hidde Ploegh HMS
Multiple myeloma (MM) affected 14,400 new individuals in the United States in 2001, with 50,000 total patients, and remains incurable despite conventional and high dose therapies. In order to overcome resistance to current therapies and improve patient outcome, novel biologically-based treatment approaches are needed which target mechanisms whereby MM cells grow and survive in the BM. Our preliminary in vitro studies show that proteasome inhibitor PS-341 mediates apoptosis of resistant MM cells, inhibits binding of MM cells to bone marrow (BM) stromal cells, and inhibits growth and survival cytokines in the BM milieu. This agent also decreases tumor cell growth and prolongs survival in a murine model, and phase II clinical trials demonstrate remarkable clinical activity, even complete responses, in patients with refractory relapsed disease. In this context, we propose to define the molecular mechanism whereby these agents mediate anti-MM activity, as well as mechanisms conferring resistance to these novel agents. To achieve this goal, we will define the role of the ubiquitin-proteasome pathway in MM pathogenesis and therapy (Specific Aim 1); define the molecular sequelae of proteasome inhibitor treatment in MM cells in vitro (Specific Aim 2); evaluate XBP-1 as a target of proteasome inhibitor therapy (Specific Aim 3); and evaluate response and resistance mechanisms to proteasome inhibitors in vivo using animal models and in derived clinical trials (Specific Aim 4). These studies will not only provide for improved use of currently available proteasome inhibitors, but will provide the framework for development of more potent and less toxic targeted therapies.
SPECIFIC AIMS:
Specific Aim 1: To define the role of the ubiquitin-proteasome pathway in MM pathogenesis and therapy
Specific Aim 2: To define the molecular sequelae of proteasome inhibition in MM cells in vitro
Specific Aim 3: To evaluate XBP-1 as a target of proteasome inhibitor therapy
Specific Aim 4: To evaluate response and resistance to proteasome inhibitors in vivo in animal models and in derived clinical trials
Project 2: Targeting Telomere Expansion Mechanisms for Myeloma Therapy
Principal Investigator:
Nikhil C. Munshi — DFCI
Co-Principal Investigator:
Ronald DePinho— DFCI
Telomeres, the specialized nucleoprotein structures at the ends of chromo—somes, protect genomic DNA from exonucleolytic degradation and end-to-end fusion. Telomere shortening occurs at each DNA replication, which if unopposed, leads to its dysfunction and genomic instability associated with transformation. Telomerase, by extending telomeric DNA, provides additional replicative capacity. Telomerase re-activation is therefore observed in most cancers and immortalized cells. In multiple myeloma (MM) cells, we have preliminary data demonstrating shortened telomeres, high levels of telomerase, an inverse relationship between telomeres and telomerase activity, and induction of apoptosis by inhibiting telomerase. Additionally we have demonstrated that IL-6 and IGF-1 maintain telomerase activity in myeloma cells via NF k B and AKT pathways. Based on the hypothesis that maintenance of telomere function is critical to myeloma cell survival, we propose to investigate mechanisms maintaining telomere function in myeloma, thereby providing the framework for targeting telomerase in novel therapeutics. To achieve this goal we will pursue the following specific aims:
SPECIFIC AIMS:
Specific Aim 1: To evaluate telomere function in MGUS, MM, and plasma cell leukemia.
Specific Aim 2: To evaluate molecular mechanisms regulating increased telomerase activity and telomere maintenance in MM.
Specific Aim 3: To evaluate telomerase/telomere-directed therapy in vitro and in derived clinical studies.
Project 3: MUC1 as a Therapeutic Target in Multiple Myeloma
Principal Investigator:
Donald Kufe — DFCI
Co-Principal Investigator:
J. Paul Eder — DFCI
The human DF3/MUC1 transmembrane protein is aberrantly expressed at high levels on the surface of multiple myeloma cells. Certain insights about the function of MUC1 have been derived from the finding that the cytoplasmic tail of MUC1 interacts with b-catenin, a signaling protein associated with the development of diverse human tumors. The interaction between MUC1 and b-catenin is regulated by receptor tyrosine kinases, members of the c-Src family and glycogen synthase kinase 3b (GSK3b). In multiple myeloma cells, stimulation with IL-7 induces interaction of MUC1 and the Lyn tyrosine kinase. Lyn phosphorylates the MUC1 cytoplasmic domain and increases binding of MUC1 to b-catenin. The functional significance of this interaction is supported by the finding that MUC1 regulates nuclear targeting of b-catenin. Our hypothesis is that MUC1 expression contributes to the malignant phenotype of multiple myeloma cells and that MUC1 represents a target for myeloma therapy.
The novel isocoumarin 2-(8-hydroxy-6-methoxy-1-oxo-1H-2-benzopyran-3-yl) proprionic acid (NM-3) has recently entered Phase I trials as an orally bioavailable inhibitor of angiogenesis. In in vitro studies of multiple myeloma cells, NM-3 disrupts binding of MUC1 to b-catenin and, as a later event, downregulates MUC1 expression. NM-3 also induces killing of multiple myeloma cells at concentrations that have been achieved clinically without toxicity. The results further demonstrate that NM-3 potentiates the effects of standard agents, such as dexamethasone, in inducing multiple myeloma cell death. We propose to define the clinical activity of NM-3 in the treatment of multiple myeloma as a single agent and in combination with agents used in the treatment of this disease. Our hypothesis is that NM-3 will induce multiple myeloma cell death, at least in part, by disrupting MUC1 signaling. This hypothesis will be addressed by studying the effects of NM-3 on MUC1 function in multiple myeloma cells in vitro and on MUC1 expression in vivo .
SPECIFIC AIMS:
Specific Aim 1: To define the functional significance of MUC1 expression in multiple myeloma cells
Specific Aim 2: To assess the role of MUC1 as a receptor in multiple myeloma cells
Specific Aim 3: To define the effects of NM-3 on MUC1 signaling and lethality in multiple myeloma cells
Specific Aim 4: To perform a Phase II pharmacokinetic/pharmacodynamic trial of NM-3 alone and in combination with standard agents for the treatment of MM.
Project 4: Novel Therapeutics Targeting Genetic Abnormalities in Multiple Myeloma
Principal Investigator:
Leif Bergsagel — Cornell
Co-Principal Investigator:
Paul Richardson — DFCI
We have shown that multiple myeloma is characterized by frequent (70%) chromosome translocations involving the immunoglobulin (Ig) genes and identified four recurrent loci that are commonly involved: 11q13, 6p21, 4p16, and 16q23. The translocations result in the juxtaposition of powerful Ig enhancers adjacent to oncogenes at these loci (cyclin D1, cyclin D3, FGFR3+MMSET, and c-maf, respectively), causing their ectopic and deregulated expression in plasma cells. Subsequent tumor progression occurs with deletion of chromosome 13, mutations of N or K ras, and secondary translocations of c-myc. The central hypothesis of this project is that genes selected by mutations are critical for the pathogenesis of myeloma and represent attractive targets for drug development. We propose to use both in vitro and in vivo model systems to validate these genes as appropriate targets for drug development, and develop assays to screen for drugs that inhibit the function of these targets. For all of this work we will first focus on FGFR3, and use it as a model to test this hypothesis. We will identify FGFR3 inhibitors by screening tyrosine kinase inhibitors entering clinical trials, and libraries of tyrosine kinase inhibitors for selective inhibition of FGFR3 auto-phosphorylation and downstream signals of FGFR3, STAT3 and MAPK phosphorylation. We will make of use of a genetically characterized panel of human myeloma cell lines, xenograft models and transgenic mouse models of plasma cell neoplasia to determine the effect of inhibiting each of these targets, and to identify surrogate markers of target inhibition. Finally we propose to introduce into clinical trials targeted agents showing promise in this pre-clinical evaluation The genetic evidence suggests that the IgH translocations are the primary event that leads to oncogenic transformation, making them a particularly attractive target. These studies will provide the framework for developing targeted therapies of myeloma, based upon the genetic abnormalities present in each patient. As with STI-571 in CML, we expect that targeting the transforming event may provide very effective therapy, and that targeting events related to progression may delay or prevent disease progression.
Specific Aim 1: To validate FGFR3 as an appropriate target for drug development using both in vitro and in vivo model systems
Specific Aim 2: To validate other genetic targets and develop surrogate markers for successful target inhibition
Specific Aim 3: Based on the preclinical data, we will carry out a clinical study targeting FGFR3 in the first year, and evaluate the anti-myeloma activity of additional agents that specifically target the pathways identified by genetic mutations in myeloma in pilot clinical trials.
Project 5: Molecular Markers Of Evolution From MGUS To Multiple Myeloma
Principal Investigator:
Rafael Fonseca — Mayo
Co-Principal Investigator:
Lynda Chin — DFCI
Monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM) are pre-malignant stages of the plasma cell (PC) neoplasm multiple myeloma (MM). Multiple chromosomal and genetic abnormalities have been described in MGUS, SMM and MM, and are thought to be important for disease pathogenesis and disease progression. We need to understand
- What genomic abnormalities place a patient with MGUS/SMM at higher risk of progression to MM?
- Which genetic loci may be lost or gained in association with evolution to MM?
Hypothesis 1: Genetic and chromosomal abnormalities present in the PCs of MGUS/SMM patients, at diagnosis, result in different risks of progression to MM. Other biologic and prognostic markers such as the PC labeling index (PCLI), bone marrow (BM) plasmacytosis, b 2 -microglobulin and concentration of the monoclonal protein are likely higher in those MGUS patients with abnormalities that put them at higher risk of disease progression.
Specific Aim 1: To test MGUS samples for the most important translocations (+14q32, t(4;14)(p16.3;q32), t(11;14)(q13;q32), t(14;16)(q32;q23), IgL- l -light chain translocations (+22q11)) and deletions (chromosome 13 monosomy, and 17p13). We will also test the samples for K/N Ras mutations, and methylation of p16.
Hypothesis 2: Chromosomal regions of recurrent gain and loss harbor genes, activation or inactivation of which is responsible, for the progression through the different stages of the PC disorders. The genome wide search for these regional alterations, and ultimately identification of the targets of such alterations will yield clues into the specific mechanisms leading to MGUS/SMM and MM.
Specific Aim 2: To generate a comprehensive genome-wide profile of regional gains and losses in MGUS and MM to identify candidate loci relevant to genesis, progression and prognosis, using array based comparative genomic hybridization.
Core 1: Administration and Communication Core
Principal Investigator:
Kenneth C. Anderson — DFCI
The purpose of the Administration and Communication Core is to assure the coordination of the Dana Farber/Harvard Cancer Center (DF/HCC) Myeloma SPORE components and to provide oversight and leadership of the scientific, administrative, and fiscal aspects of the SPORE. The SPORE Director will oversee the administrative coordination of the various clinical and laboratory studies outlined in this Program Project. He will integrate scientific and clinical efforts within and between Projects and assure the translation of laboratory findings to the bedside; and conversely, the initiation of laboratory studies stemming from clinical observations. This Core will also facilitate exchange of information among the SPORE members, as well as the internal and external advisory committees. It provides clinical research nursing support for the proposed clinical trials. In addition, a clinical study coordinator will assure appropriate sample acquisition and trafficking. The grants administrator will allocate resources in a timely and integrated fashion to facilitate successful completion of the proposed studies.
- Monitor research progress and plan for the future;
- Foster collaborative research within the SPORE and between SPOREs;
- Integrate the Myeloma SPORE into the DF/HCC structure;
- Provide necessary resources and fiscal oversight;
- Promote rapid dissemination of significant research finding
Core 2: Tissue Core
Principal Investigator:
Phil Greipp — Mayo
The goals of the Core Laboratory and Tissue Bank are to aid SPORE investigators: 1) to identify tumor biologic and molecular genetic correlates of response, progression, and survival in the context of the SPORE proposal by performing appropriate assays; 2) to maintain a searchable database of clinical and laboratory data for use by SPORE investigators; and 3) to provide a resource for banked specimens for future studies. Core Laboratory functions are designed to increase the power of the individual projects to detect biologic differences among patients entering the SPORE, and to be flexible enough to address individual investigator needs at different times. As in similar Mayo Clinic based myeloma core facilities this Core receives, processes, stores, archives and distributes serum, bone marrow cells, DNA, and RNA only for investigators participating in this SPORE. Samples from the SPORE will be collected, tested, and archived separately from other Core resources such as the ECOG or the Mayo Program Project reference laboratories. This Core provides an independent, current, and quickly searchable database of clinical and laboratory results and archived biospecimens. Bone marrow cells and sera collected at SPORE centers are sent to the Core Laboratory using kits with pre-paid mailers including complete instructions and materials. Data forms completed at the referring institutions at the time of collection of the samples are entered into a common clinical, laboratory, and archival database similar to that used in the ECOG Cooperative Group Myeloma Reference Laboratory and the Mayo Program Project grant. Follow up data is obtained and entered as required. All data will then be stored in a common database accessible by the Statistics center. All patients will have central laboratory measurement of the plasma cell labeling index and other key biological variables requested by SPORE investigators. Unused specimen including sorted cells, DNA, RNA and cytospin slides will be stored for future use on all patients. Subsequent use of banked samples beyond that specified in this SPORE proposal will be provided only with agreement from Principal investigators and the SPORE Principal Investigator. This Core is lead by investigators experienced in managing Core laboratories in a multi-institutional cooperative setting.
Core 3: Biostatistics Core
Principal Investigator:
Edie Weller — DFCI
The purpose of the Biostatistics Core 4 is to provide the following services that will be utilized by the research projects and the clinical studies in the SPORE.
Specific Aim 1. To provide biostatistical collaboration for clinical research protocols. This includes all aspects of design, conduct, analysis and reporting of the clinical protocols; and direction of data processing and quality control assurance.
Specific Aim 2. To provide biostatistical collaboration for animal and laboratory research studies. This includes all aspects of design, conduct, analysis and reporting of such studies, including the coordination of laboratory results with parameters and outcomes from the clinical studies.
Specific Aim 3. To supervise the data processing and data quality assurance, and to coordinate the efforts of the data coordinator and those of the data analyst in maintaining an accurate computerized database.
Specific Aim 4. To provide computing resources for data processing, forms design, and statistical analysis, standardized reporting, and quality control.
DEVELOPMENTAL PROJECTS
The objectives of the Developmental Projects Program are to provide a continuous flow of new ideas and projects to stimulate myeloma research in the context of the Myeloma SPORE. It encourages new research directions and methodologies and facilitates collaborations. By providing initial support to pilot projects, it will foster the development of new translational projects. It also allows the Myeloma SPORE to have participation and recruitment of investigators not only from the DF/HCC, but also including outside institutions such as Mayo Clinic, University of Arizona Medical School, and Cornell University Weil Medical School. This Program will rely on scientific and programmatic review by a Developmental Research Committee, which will assure selection of the most promising, highest quality, projects with high likelihood of translational impact. The projects selected for the first year are
- Multi-targeted therapeutic interventions on heat shock responses in myeloma - Nicholas Mitsiades, M.D. Dana Farber Cancer Institute
- Homologous recombination, DNA instability, and multiple myeloma: a promosing new target for therapy and prevention. - Masood Shammas, Ph.D. Harvard Medical School
CAREER DEVELOPMENT AWARDS
The investigators assembled in the DF/HCC Myeloma SPORE have a substantial long-term record in mentorship and development of junior faculty working in translational myeloma research. The goal of the Career Development Program of our Myeloma SPORE is to build upon this record and establish a formal process for the identification, selection, funding, and mentoring of individuals pursuing careers in the study of the basic and clinical aspects of myeloma. These awards will facilitate the development of physicians, physician scientists, clinical investigators, and scientists in training within the Myeloma SPORE Program towards faculty status. Thus, candidates will be junior faculty or fellows and postdoctoral fellows within the various training programs across DF/HCC and participating institutions, including the campuses at Harvard, Mayo Clinic, University of Arizona, and Cornell University. It is our goal to attract, mentor, and assure the success of several candidates within the timeframe of this SPORE. Success will be defined as the development of physician/scientists in training towards careers as independent investigators. The Four career development awards in the first year are:
- Investigation of the celllar- and biodistribution of proteasome inhibitors, effects on proteasomal activity and composition in vivo. - Huib Ovaa, Ph.D. Harvard Medical School.
- Generation of a comprehensive genome-wide profile of regional gains and losses in human mutiple myeloma to identify new candidate loci relevant to genesis, progression and prognosis, using array based comparative genomic hybridization (aCGH) - Ruben Carrasco, M.D. Brigham and Women's Hospital/Harvard Medical School.
- Multiple Myeloma and the unfolded protein response. Ann-Hwee Lee, Ph.D. Harvard School of Public Health.
- Preclinica;l development of therapeutic strategies to enhance the activity of conventional or investigational anti-myeloma agenetsa by inhibition of the IGFs/IRF-1R axis. Constatine Mitsiades, M.D.,Ph.D. Dana Farber Cancer Institute.
