Grant: 6550-18 | Translational Research Program (TRP):
Location:Dana-Farber Cancer Institute, Boston, Massachusetts 02215
Project Title: Prioritizing The In Vivo Therapeutic Relevance Of "myeloma-selective" Essential GenesProject Summary:
Multiple myeloma (MM) remains incurable because its malignant cells manage to eventually escape from even the most potent combinations of new therapies for this disease. Despite years of extensive research, the MM field has not yet comprehensively characterized which genes are essential for MM cells to survive and grow within the supportive microenvironmental niche of the bone marrow (BM). We applied, in our lab, the exciting recent technology of CRISPR/Cas9 genome editing to simultaneously investigate the function of thousands of genes. We identified in our studies a select set of genes that are considerably more important for survival and growth of MM cells in the laboratory, compared to tumor cells from other blood cancers or from solid tumors. Some of these "MM-selective" essential genes have known roles in the biology of MM pathophysiology, but several others have not been previously considered major therapeutic targets. This project will validate the therapeutic relevance of these "MM-selective" essential genes using a model in which biocompatible scaffolds are implanted into mice and are engineered, with the use of human stromal cells from the BM, to resemble the properties and local conditions of the BM in patients with MM. Building on our experience with the CRISPR/Cas9 system both in the lab and in mouse studies, we will examine in this "humanized" BM-like scaffold model which of our candidate genes remain essential for MM cells in this context and therefore represent promising therapeutic targets. Investigational drug-like small molecules are available for only a few of these "MM-selective" essential genes, while most of the latter are deemed currently "undruggable". We observed though that specific small molecule inhibitors against other targets involved in distinct components of the regulation of gene expression can significantly down-regulate the transcript levels of several of the most prominent "currently undruggable" MM-selective essential genes in MM cells, and indirectly can disrupt the molecular network of these genes. We will therefore examine if small-molecule inhibitors targeting MM-essential genes, directly or indirectly, exhibit activity against MM cells in the "humanized" BM-like scaffold-based model. By combining the powerful CRISPR technology with in vivo models simulating the human BM, and pharmacological agents with direct or indirect activity against individual or multiple candidate MM-selective essential genes, we hope to determine the therapeutic relevance of these gene and provide a framework to guide the translation of inhibitors for these targets towards the clinic for MM patients.
Grant: 1349-18 | Career Development Program (CDP):
Location:Washington University School of Medicine in St. Louis, St. Louis, Missouri 63112-1408
Project Title: Protection Of Proliferating B Lymphocytes From Transformation By A C-MYC-induced Tumor Suppressive ProgramProject Summary:
Lymphomas and leukemias are caused by uncontrolled proliferation of lymphocytes due to accumulating errors in the genome. However, cell proliferation is also an important biological activity across many different tissues and cell types. Specifically, proliferation of lymphocytes is essential for the immune responses that protect individuals from invading pathogens. Normal lymphocytes are able to proliferate even quicker than cancer cells in response to infection for extended periods of time. In the course of normal immune system development, lymphocytes also mutate their own genome to establish better immunity to infection. However, it is unknown how lymphocytes undergo normal proliferation and mutation events without increasing the risk of cancers. We hypothesize that lymphocytes engage an unidentified, unique mechanism to minimize the risk of cancers while facing the demand to proliferate and mutate their genome during their normal functional activities. To explore this hypothesis, we have studied sets of genes that are activated by a protein named c-Myc, which has been causally linked to many cancers in humans and other organisms. Since c-Myc is also essential for lymphocyte proliferation in response to infection, c-Myc may activate an unidentified pathway that is required to protect normally proliferating lymphocytes from becoming cancerous. Accordingly, mutations of genes in the pathway may increase the risk of lymphomas or leukemias. Indeed, we have identified a gene that is activated by c-Myc in proliferating lymphocytes and is necessary to suppress blood cancer development in an animal model.Moreover, a few mutations of this gene have been found in leukemic cells from human patients.Starting with these discoveries, we will define the molecular mechanisms by which this c-Myc initiated regulatory pathway suppresses leukemia and lymphoma. The knowledge from this study will contribute to the development of improved strategies for the risk assessment and diagnosis of leukemias and lymphomas and also to inventing new therapeutic approaches.
Grant: 3378-18 | Career Development Program (CDP):
Location:Sloan Kettering Institute for Cancer Research, New York, New York 10087
Project Title: Optimizing CAR T Cell Therapy For Multiple MyelomaProject Summary:
While advances have recently been made in the management of multiple myeloma (MM), MM is still considered incurable, with most patients dying from their disease. Thus, there is a critical need to identify and evaluate new therapeutic modalities that may induce durable remissions or cures.
T-cells keep the body healthy by killing cells infected with bacteria or viruses. Chimeric antigen receptor (CAR) T-cell therapy involves collecting a patient’s T-cells and genetically reprogramming them in the lab to recognize and kill cancer cells instead of infected cells. The cells are then infused back to the same patient where they serve as a living drug. In relapsed or refractory acute lymphoblastic leukemia (ALL) this strategy led to dramatic complete responses in approximately 80% of patients treated.
To extend the effectiveness of this therapy to MM, we and others developed MM-specific CARs. Specifically, we screened tens of billions of human antibody fragments and identified a handful that bound to a specific protein target unique to MM cells. We then used the DNA encoding these ‘hits’ to make CARs. Subsequently, we compared how T-cells modified to contain these different CARs preformed in a variety of tests in cell cultures and in mice. We found one CAR, which conveyed very favorable properties on T-cells, to test in patients. We recently began treating MM patients on the clinical trial to test this novel CAR T-cell therapy.
Based on our experience with CAR T-cell therapy for ALL, we hypothesize that patients may develop resistance to our therapeutic approach. Potential sources of resistance may include loss of our target protein on the MM cells and/or suppression by other cells in the environment in which the MM cells reside. We will evaluate the effectiveness, including resistance mechanisms, of our CAR T-cell therapy by analyzing samples obtained through clinical trial. We are also developing “armored CARs,” which express a second protein that may alter the environment in a way that enhances anti-tumor immunity. Additionally, we will study patient samples in a unique mouse model that can support both MM tumor cells and cells found in the environment in which MM cells normally reside in patients. This model will be used to investigate whether our next generation CAR vectors, including CAR T-cells targeting MM-specific proteins (our armored CAR) may enhance long term anti-MM immunity.
Grant: 5469-18 | Career Development Program (CDP):
Location:Joan & Sanford I. Weill Medical College of Cornell University, New York, New York 10022
Project Title: Mutations Disrupting Gene Enhancer Epigenetic Complexes As Drivers Of Lymphomagenesis.Project Summary:
Lymphoma is a form of cancer that affects immune cells called lymphocytes, a type of white blood cell. There are many subtypes and maturation stages of lymphocytes and, therefore, many kinds of lymphomas. Follicular lymphomas (FL) develop from B lymphocytes (B-cells) and are the second most common subtype of non-Hodgkin lymphoma. Clinically, FL is an indolent lymphoma, characterized by slow progression and relatively high overall survival rate. However, FL is generally considered incurable and roughly half of all FL patients eventually progress into a more aggressive lymphoma with very poor prognoses.
During the normal immune response, B-cells undergo a stage where they grow much faster than usual and tolerate changes in their genetic material, allowing them to develop into fine-tuned functional immune effectors. Three proteins, called BCL6, SMRT and NCoR, work together to drive this process of terminal differentiation in a tightly controlled manner. Any alterations, such as those occurring in FL, can cause cells to become unresponsive to signals cueing exit from this program. B-cells thus become “frozen” at this stage of development; they are unable to complete their normal program and fail to die when instructed to, collecting in the lymph nodes and forming tumors.
Recently, through analyzing the genomes of FL patients, our laboratory has identified several mutations in a gene called TBL1XR1. Our preliminary data suggest that mutations in TBL1XR1 may result in increased stability of SMRT and NCoR. In this study, we plan to investigate whether the increased stability of SMRT and NCoR proteins by TBL1XR1 mutants can cause enhanced BCL6 activity and in turn lead to the development of FL. Unveiling the mechanisms by which B-cells cease to respond to their normal regulatory signals will profoundly contribute to our understanding of FL. In addition, our study will explore whether TBL1XR1 mutations can predict which FL patients have a BCL6-dependent disease and thereby help identify patients who could benefit from existing drugs targeting this protein.
Grant: 8011-18 | Screen to Lead Program (SLP):
Location:The Trustees of Columbia University in the City of New York, Columbia University Medical Center, New York, New York 10027
Project Title: Development Of NT5C2 Inhibitors For Treatment Of Relapsed Refractory ALLProject Summary:
Despite intensive chemotherapy, 20% of pediatric and over 50% of adult acute lymphoblastic leukemia (ALL) patients fail to achieve a complete remission or relapse after intensified chemotherapy, making relapse and resistance to therapy the most significant challenge in the treatment of this disease. This project seeks to develop highly active and specific inhibitors of NT5C2, a protein activated by mutations in relapsed leukemia cases. NT5C2 is directly responsible for chemotherapy resistance, making NT5C2 inhibitor an attractive strategy for the treatment of relapsed and refractory ALL patients.
Grant: 5470-18 | Career Development Program (CDP):
Location:Board of Trustees of the Leland Stanford Junior University, San Francisco, California 94144-4253
Project Title: Dissecting The Topological Consequences Of Mutations In The Cohesin Complex And Their Contribution To Human Leukemia Initiation And Progression.Project Summary:
The human genome is exquisitely organized, packing five feet of DNA into a microscopic nucleus. This level of compaction requires an equally impressive level of organization. Not only does the DNA have to fit into such a tiny space; the genes required for the cell’s function must be properly expressed with high fidelity. To accomplish this seemingly insurmountable task, the genome is organized into a cascading series of loops whose formation is mediated by a multi-protein complex called the cohesin complex. This complex forms a ring-like structure that holds the ends of each loop together much like the clasp of a necklace. Recently, mutations in the cohesin complex have been identified as major drivers of acute myeloid leukemia initiation and progression. With such an important basic cellular function as the organization of DNA, it remains of vital importance to understand how mutations in the cohesin complex mechanistically cause leukemia and to identify avenues for therapeutic intervention.
Of the core components of the cohesin complex, the STAG2 gene is the most frequently mutated and is often lost altogether. Using state-of-the-art technologies, we will study how the composition and function of the cohesin complex changes in the context of a loss of the STAG2 protein. These technologies include the ability to map the three-dimensional organization of the genome, enabling us to study how the DNA loops that are orchestrated by the cohesin complex are disrupted in AML. These molecular and mechanistic studies will be accompanied by genetic and pharmacologic screening to identify putative therapeutic targets. Ultimately, this research will bring us closer to understanding how acute myeloid leukemia develops and how dysregulation of genome organization contributes to this process. We hope that our work will identify promising drug targets and facilitate the treatment of this otherwise deadly disease.
Grant: R6509-18 | Translational Research Program (TRP):
Location:University Health Network, Toronto, Ontario M5G 1Z5
Project Title: Phase I Study Of Allogeneic Double Negative T Cells In Patients With High Risk AMLProject Summary:
Acute Myeloid Leukemia (AML) is a cancer affecting the bone marrow that requires intensive chemotherapy for disease control. This may be associated with significant toxicity. Treatment for AML aims to destroy the leukemia cells and allow the bone marrow to work normally again. Chemotherapies help most AML patients to achieve a state of remission in which the leukemic cells have fallen to a very low level and normal blood cell production has returned. However, despite decades of using chemotherapy to treat AML patients, there continues to be a very high chance of the disease coming back as the cancer cells develop resistance to chemotherapies. Once the disease comes back, few treatment options are available to patients. Consequently, overall survival is about 40% for those <50 years of age and is significantly worse for older patients. Thus, new treatments that are effective at targeting chemotherapy-resistant AML with low toxicity are needed for this unmet challenge.
Double negative T cells (DNTs) are a type of white blood cell in the blood that has the ability to selectively kill cancer cells while sparing normal cells and tissues. In peripheral blood, DNTs are quite rare. To be of value in the treatment of cancer it is necessary to have large numbers of these cells. We have developed methods in the laboratory to grow DNTs from healthy volunteers to large numbers and have shown that these DNTs have potent anti-leukemia effect without detectable toxicity on normal cells and tissues in animal models. In this first-in-man clinical trial, DNTs will be obtained from healthy donors and expanded (increased in numbers) in the laboratory, in order to enhance their tumor destroying potential. In addition, the most potent DNT donor will be selected for each patient by screening before his/her DNT treatment. DNTs will be administered at 3 different dose levels (different numbers of DNT cells) into 3 different groups of patients. After infusion of the first (lowest cell number) dose level, patients will be monitored to make sure they do not experience any severe adverse side effects. Only after the confirmation of the safety of the previous dose, the next group of patients will be given the next dose (higher cell number). In addition to determining the safety of DNT treatment, we will take blood samples from the treated patients at several time points after infusion to assess how long the DNT cells are detectable in the patients, which will provide guidance on the optimal number of injections and the time between each injection required to achieve the best results. In addition, we will determine if infusion of DNT cells changed the amount of time for the patients’ immune cells to recover from chemotherapy. Furthermore, whether DNT treatment will decrease AML cells and reduce the rate of the disease returning will be monitored. The results from this study will indicate whether DNTs can be safely used as a new treatment for AML to save patient lives.
Grant: 6555-18 | Translational Research Program (TRP):
Location:The University of Texas MD Anderson Cancer Center, Houston, Texas 77210-4266
Project Title: Immunotherapy For Multiple Myeloma Using Off-the-Shelf Cord Blood Derived Natural Killer CellsProject Summary:
Multiple myeloma (MM) is caused by the malignant transformation of plasma cells. High dose chemotherapy followed by stem cell transplantation from a matched healthy donor (allogeneic stem cell transplantation) offers a potentially curative treatment for advanced cases of this disease. Unfortunately, only about 25% of MM patients can expect to benefit from this approach, mainly because of the high risk of infection and other toxicities associated with allogeneic stem cell transplantation, as well as the high relapse hazard that defines resistant MM. We propose to take advantage of an exciting new weapon, cancer immunotherapy, to combat high-risk MM without the risks associated with allogeneic stem cell transplantation. Natural killer (NK) cells, isolated either from the patients themselves or from normal adult donors, are being used increasingly because of their potent anti-cancer effects. We have identified umbilical cord blood as an ideal source of NK cells, as the cells are already frozen and ready for use, without the need for collection from an adult donor. The aim of this research is to harness NK cells to destroy MM cells. Recent progress in our laboratories has shown that NK cells can be isolated from cord blood and expanded outside the body to large enough numbers to be used in adult patients, where they are expected to kill MM cells efficiently without introducing major toxic effects. This NK cell activity, when combined with high dose chemotherapy, a novel drug called lenalidomide (which promotes the anti-myeloma effects of NK cells), and elotuzumab (an antibody that targets a protein on the surface of myeloma cells) could establish an entirely new strategy of effective immunotherapy for MM that may eliminate the need for allogeneic stem cell transplantation in this disease. Finally, we propose to boost the anti-myeloma potential of NK cells by engineering them to increase their ability to recognize and kill MM cells using a mouse model of the disease. If our attempt at specific targeting is successful in the laboratory, we will move this strategy into clinical testing as rapidly as possible.
Grant: 6544-18 | Translational Research Program (TRP):
Location:Mayo Clinic, Rochester, Minneapolis, Minnesota 55486-0334
Project Title: Modulating Immune Function In Peripheral T-cell Lymphoma (PTCL)Project Summary:
Patients with peripheral T- cell lymphoma (PTCL) constitute approximately 12-15% of all lymphoma cases and PTCL patients typically have a poor outcome. Patients with PTCL typically respond to initial combination chemotherapy, but most patients subsequently progress and require additional therapy. Treatments such as romidepsin and belinostat have been approved for patients with PTCL, but their efficacy has been limited. There is therefore clearly a need for additional new therapies to treat patients with PTCL.
Multiple mechanisms account for the lack of durable benefit with current treatment, but the inability of tumor-specific immune cells to eradicate the lymphoma is a central issue. A novel mechanism used by cancer cells to inhibit the immune response, however, is overexpression of programmed death ligand 1 or 2 (PD-L1 or PD-L2). These proteins interact with the programmed death-1 (PD-1) receptor expressed on intratumoral T-cells and provide an inhibitory signal thereby suppressing anti-tumor immunity. In previous work, we have shown that PD-1/PD-1 ligand interactions are critically important in PTCL and have found that PD-1 is expressed both on malignant cells and in the tumor microenvironment. Similarly, we have found that PD-L1 is highly expressed by malignant T-cells and intratumoral monocytes, and have shown that tumor-associated PD-L1 suppresses T-cell immunity.
Monoclonal antibodies that block PD-1 signaling can prevent T-cell suppression and promote an anti-lymphoma immune response. Antibodies, directed against PD-1 or PD-L1, are currently being tested in clinical trials and have shown remarkable efficacy in some hematologic malignancies. Despite broad clinical use of PD-1 directed therapy, very little is known about the immunology of PD-1 signaling in hematological malignancies. In PTCL in particular, PD-1/PD-L1/2 signaling is complicated by the fact that the receptor and ligands can both be expressed on the cancer cell. Furthermore, initial clinical trials have included very few patients with PTCL; however, in 5 patients receiving nivolumab, an antibody that blocks PD-1 signaling, 2 patients had an objective response, suggesting possible efficacy of PD-1 blockade in PTCL.
In this application, therefore, we propose to 1) measure the immune consequences of inhibiting PD-1/PD-1 ligand signaling, 2) determine the clinical efficacy of PD-1 blockade alone, and in combination with romidepsin, in patients with relapsed and refractory PTCL, and 3) determine the immunological predictors of clinical response to PD-1 therapy in patients with PTCL. In the proposed aims, we will determine how well PD-1 blockade works in PTCL, the specific mechanism of action of PD-1 blockade in PTCL, and in which subgroup of PTCL patients PD-1 blockade is predicted to have clinical benefit. Successful completion of this project is likely to have a major impact on clinical practice by potentially leading to an effective new therapy for patients with PTCL.
Grant: 6539-18 | Translational Research Program (TRP):
Location:Board of Trustees of the Leland Stanford Junior University, San Francisco, California 94144-4253
Project Title: Applying An Innovative Microscopy Platform To Study Lymphoma In The Context Of A New Clinical TrialProject Summary:
For decades, cancer treatment has relied on toxic chemotherapy to kill cancer cells, which also kills normal cells in the body and causes severe side effects. We have entered a new era of cancer therapies in which we harness the precision of the body’s immune system to seek and destroy each cancer cell, akin to how it rids the body of infections. However, this result currently remains out of reach for the vast majority of cancer patients. There are two main hurdles to overcome. First, we need better strategies that work in a larger proportion of patients. Second, and related, we need a better understanding of the ingredients necessary to produce a powerful immune response against the cancer.
To address the first challenge, our group has focused on a strategy called in situ vaccination, which involves injecting immune stimulants directly into a tumor to activate the body’s immune cells, which can then recognize and kill tumor cells anywhere in the body. These cells will also persist as memory cells to prevent the cancer from returning, similar to how a vaccine can prevent an infectious disease from taking root. The trick is to find the right combination of immune stimulants to make this work. In multiple mouse models of cancer, a small molecule called SD-101 and an antibody drug that recognizes the OX40 protein on immune cells combine to produce this exact result. SD-101 is a synthetic molecule that mimics bacterial DNA, thus acting as a danger signal that activates the immune system. We have found that this activation increases the amount of OX40 protein on T cells in the tumor, the immune cells responsible for recognizing and killing tumor cells. The anti-OX40 antibody boosts the activation of these T cells while also suppressing regulatory cells that dampen the immune response. In mice growing two tumors under their skin, injecting these two agents directly into just one tumor results in both tumors disappearing. We now propose to study this novel approach in patients with lymphoma in a clinical trial.
With regard to the second challenge, we have struggled to understand how cancer cells and immune cells interact because we lack techniques to study these processes broadly in small patient samples. Thus, we are collaborating to pilot a novel microscopy platform that simultaneously identifies a diverse array of cell types and proteins in a patient’s cancer, while preserving the tissue architecture, something that has not previously been possible. We propose to use this technique to study samples from patients as they undergo our combination treatment, to provide an unprecedented look at the precise interactions between the immune system and cancers. Armed with this knowledge, we hope to one day bring the power of immunotherapy to all cancer patients.