Grant: 6557-18 | Translational Research Program (TRP):
Location:University of Perugia. Division of Hematology and Clinical Immunology, Department of Clinical and Experimental Medicine, Perugia, 06132
Project Title: HAIRY CELL LEUKEMIA (HCL): A CHEMOTHERAPY-FREE TREATMENT STRATEGY CENTERED AROUND BRAF INHIBITIONProject Summary:
We discovered a mutation of the BRAF gene as the genetic cause of Hairy Cell Leukemia (ref. 1). We also found in the laboratory that drugs designed to block mutated BRAF kill patients' hairy cells, while sparing cells laking this mutation (ref. 2), including normal cells. Thus, inhibiting mutated BRAF could represent a new approach to the treatment of HCL, which is currently based on myelotoxic chemotherapeutics (purine analogs) killing not only leukemic cells but also normal bone marrow (i.e., myeloid) cells.
Indeed, we showed that a brief treatment by mouth with the BRAF inhibitor vemurafenib proved highly active in HCL patients relapsed after, or refractory to, chemotherapy, without the toxic effects of the latter (ref. 3). However, the disease tends to come back some time after stopping vemurafenib intake due to the persistence, at the end of treatment, of some residual hairy cells in the bone marrow. These residual hairy cells represent the reservoir for subsequent leukemia regrowth and, in about half of patients, they manage in some way to bypass BRAF and reactivate MEK, which is the target of BRAF and which should be deactivated when BRAF is inhibited by vemurafenib (ref. 3).
This project aims at improving on these results by rationally adding to vemurafenib other "intelligent", non-toxic drugs, in order to eradicate all leukemic cells and therefore achieve lont-term complete remssions. In particular, in a set of patients we will combine vemurafenib with cobimetinib, an inhibitor of MEK that is also taken by mouth, in order to impede MEK reactivation during therapy with vemurafenib. In another set of patients, we will combine vemurafenib with obinutuzumab, a monoclonal antibody given intravenously, that recognizes a molecular marker (CD20) on the surface of all hairy cells (i.e., irrespective of whether or not they managed to reactivate MEK); in so doing, obinutuzumab is expected to stimulate the immune system to attack leukemic cells from the outside, while vemurafenib is concurrently blocking mutated BRAF inside them (two-pronged strategy). Finally, if patients still relapse after either of these two drug combinations (vemurafenib plus cobimetinib; vemurafenib plus obinutuzumab), we will use all these three drugs together to try to get these patients in remission again.
We will accomplish this goal through a clinical trial that will be conducted in multiple centers throughout Italy, and that will be open to patients that have relapsed/refractory HCL or that are unfit for chemotherapy (e.g., due to old age, frail medical conditions, or ongoing infections which could be worsened by chemotherapy).
This project will help to clarify the most attractive combination of "intelligent" drugs to be used in relapsed/refractory HCL patients, and to be selected for potentially challenging purine analogs in newly diagnosed patient with this leukemia, in the prospect of a chemotherapy-free therapeutic strategy for all HCL patients.
Grant: 6536-18 | Translational Research Program (TRP):
Location:The University of North Carolina at Chapel Hill, Atlanta, Georgia 30384-2420
Project Title: Exploiting The Inducible Caspase9 To Pharmacologically Modulate CD19.CAR-T Cell Function In VivoProject Summary:
The administration of a subset of human immune cells cultured in the laboratory and known as T lymphocytes that have been engineered to express a chimeric molecule that recognizes leukemic cells (CD19.CAR-Ts) has shown remarkable antitumor effects in patients with a blood tumor known as acute lymphoblastic leukemia (ALL). We propose here to improve this therapy by reducing its potentially lethal side effects that are caused by the extreme expansion of these cells once they encounter tumor cells in patients. These side effects are known as 1) cytokine release syndrome, with symptoms varying from flu-like symptoms to more severe side effects such as cardiac arrest, renal failure and ultimately death; 2) seizure and coma that are often life threatening; 3) life-long inability to recover the normal cells of the blood that protect from infections. We have now developed and validated in the laboratory a strategy that allows a precise regulation of the expansion of the T cells in patients, by using an absolutely safe drug that only works on the T cells infused into the patients, thus significantly controlling the cytokine release syndrome. In addition, a slightly higher dose of the same drug can be used to completely eliminate the T cells infused when the patient has achieved a durable elimination of the tumor cells. We now propose to use this strategy to in a phase I clinical trial (LCCC1541-ATL) to determine whether receiving iC9-CAR-Ts cells is safe and tolerable in adult and pediatric subjects with relapsed/refractory ALL and to determine our capacity to control the expansion of the T cells once infused into the patients. We have written the clinical trial, which has received approvals from the various regulatory agencies and validated the production of cells in our specialized manufacturing facility.
Grant: 6531-18 | Translational Research Program (TRP):
Location:The Trustees of Columbia University in the City of New York, Columbia University Medical Center, New York, New York 10027
Project Title: New Therapies In Relapsed ALLProject Summary:
Acute lymphoblastic leukemia (ALL) is an aggressive hematological tumor resulting from the malignant transformation of early lymphoid progenitor cells. Despite intensive chemotherapy, 20% of pediatric and over 50% of adult ALL patients fail to achieve a complete remission or relapse after intensified chemotherapy. Importantly, relapsed ALL is frequently associated with chemotherapy resistance and, despite salvage therapy with intensified treatment, cure rates are still unsatisfactory low. Here we propose to explore the therapeutic role of inosine monophosphate dehydrogenase (IMPDH) inhibitors in the treatment of relapsed and chemotherapy resistant ALL. Specifically we will ask: what is the specific role and mechanisms that mediate the antileukemic effects of IMPDH inhibitors in relapsed leukemia?; What patients would benefit best from treatment with these drugs?; and finally, what drug combinations may enhance the antitumor activity of these therapeutics? These studies will be instrumental for the clinical development of IMPDH inhibitors in the treatment of high risk leukemia.
Grant: 1346-18 | Career Development Program (CDP):
Location:University of Cincinnati, Cincinnati, Ohio 45221-0222
Project Title: The Oncogenic Role And Underlying Mechanism Of TET1 In Acute Myeloid LeukemiaProject Summary:
Acute myeloid leukemia (AML) is one of the most common and fatal forms of hematopoietic malignancies. Thus, it is urgent to better understand the mechanisms underlying the pathogenesis of AML, and on the basis of such understanding, to develop novel therapies with higher efficacy and minimal side effects to treat AML. The properties of cancer are often determined by the proteins that are expressed from information provided by the genes in the cell. Expression of information from genes is regulated in part by chemical modifications of the DNA in the gene, in a process called “epigenetic regulation.” One such modification is called methylation. A family of proteins involved in epigenetic regulation are the 3 TET proteins, which ultimately affect the methylation status of critical genes. TET proteins are traditionally thought to be negative regulators of tumor growth. However, in contrast to this tumor suppressive role, we recently reported that TET1 is highly expressed in certain subtypes of AML, suggesting an opposite role for TET1, the promotion of tumor growth.
We are currently studying the mechanisms by which TET1 promotes tumorigenesis and how we might use this information to develop a novel approach to treat AML. Some central questions are to understand how tumors develop, how they are maintained after they develop, and how the leukemia stem cells provide a reservoir for continued tumor development in a patient. Therefore, we seek to understand the role of TET1 in both the development and maintenance of the AML types that overexpress TET1, and the role of TET1 in the leukemic stem cells. In addition, we are studying the critical target genes that are affected by TET1, which will provide further insight into the role of TET1 in AML. Lastly, we are examining ways to therapeutically target TET1 using mouse models of AML. The success of our studies will provide novel insights into our understanding of the critical role of TET1 in AML and may also lead to the development of novel and more effective therapeutic approaches to treat the AML.
Grant: 5466-18 | Career Development Program (CDP):
Location:The Wistar Institute, Philadelphia, Pennsylvania 19104
Project Title: The Role Of EBNA1 In Epigenetic Regulation Of Gene Expression And EBV LatencyProject Summary:
Epstein-Barr virus (EBV) is a human tumor virus responsible for over 200,000 cancers per year, including multiple blood cancers such as Burkitt’s lymphoma, Hodgkin’s lymphoma, and NK/T cell lymphoma. Like all herpesviruses, EBV can develop a long-term, largely dormant phase called latency, with only occasional reactivation (called the lytic phase). Unlike most other viruses,however, EBV-associated pathogenesis depends on viral latency, rather than an active, lytic infection. During latency, only a handful of viral proteins are expressed, and among these only EBV nuclear antigen (EBNA)-1 is expressed across all forms of EBV-associated cancers. Although it is known that EBNA1 plays a central role in regulating both viral and host gene expression, the mechanisms associated with this regulation remain incompletely understood. Interestingly,epigenetic regulation, or mechanisms of altering gene expression beyond changes to the genetic code, has been shown to play a significant role in cancer development and plays a role in maintaining EBV latency. While EBNA1 is vital in establishing EBV latency and maintaining the latent viral genome, the role that EBNA1 plays in regulating host gene expression and cancer cell development remains unclear. To better understand EBNA1, we will use various approaches to investigate the role of EBNA1 in regulating gene expression on an epigenetic level, where the proteins bound to DNA are modified. We have previously demonstrated that EBNA1 is a direct regulator of genes involved in cell proliferation and survival, and our current studies will expand our knowledge of the mechanism of this regulation and the identification of additional direct targets of EBNA1. A better understanding of EBNA1-mediated gene regulation will give us the opportunity to investigate new mechanisms for inhibiting the function of EBNA1 and validate the potential of EBNA1 as a therapeutic target.
Grant: 3377-18 | Career Development Program (CDP):
Location:Sloan Kettering Institute for Cancer Research, New York, New York 10087
Project Title: Understanding The Effects Of Leukemia-Associated Mutations In Spliceosomal Proteins On Chromatin StateProject Summary:
In the past few years, genetic analysis of leukemias has identified frequent mutations in a class of genes that encodes for proteins participating in a process called RNA splicing. Mutations in RNA splicing factors are now known to be the most common type of mutation in patients with myelodysplastic syndromes (MDS) and related myeloid leukemias as well as chronic lymphocytic leukemia (CLL). These discoveries have resulted in intense efforts to understand how mutations in RNA splicing factors promote the development of leukemia.
RNA splicing is the process whereby genetic information is read from DNA and used to make proteins. Currently, most efforts to study RNA splicing factor mutations have focused on the effects these mutations have on the process of RNA splicing itself. RNA splicing factors, however, are known to play additional roles not directly related to splicing. In accordance with this, we have identified a unique effect of RNA splicing factor mutations on the epigenome. The epigenome refers to chemical changes on chromatin, which are structures in the cell made up of DNA and the proteins surrounding DNA. These chemical changes regulate which genes are expressed from DNA and when they can be turned on and turned off. Based on our preliminary results, we believe that one of the main ways that RNA splicing factor mutations cause leukemia is by altering the epigenome. We have shown that one of the most commonly mutated splicing genes, SF3B1, produces a protein that binds to some parts of chromatin. However, the extent of this binding to different chromatin components and the role that this binding plays in altering the epigenome needs clarification. We are now studying this relationship between RNA splicing factor mutations and the epigenome in more precise detail. Our longer term goal is to utilize this information to develop new therapeutic approaches for leukemia cells carrying RNA splicing factor mutations.
Grant: 2318-18 | Career Development Program (CDP):
Location:Charlotte Mecklenburg Hospital Authority d/b/a Carolinas HealthCare System, Charlotte, North Carolina 28203
Project Title: Optimizing Risk And Response Adaptive Strategies Using Immunotherapy In Multiple MyelomaProject Summary:
Despite the more than three-fold improvement in survival outcomes over the last 15 years, multiple myeloma (MM) remains an incurable disease. There is growing recognition that MM disease biology is complex and that personalized treatment strategies need to be developed for different MM patients. However, the current MM treatment paradigm is largely based on a patient’s eligibility for a stem cell transplant. Thus, an incredibly heterogeneous disease is being treated in a ‘one-size-fits-all’ way that translates into broad variability in patient outcomes. As anti-myeloma therapeutics continue to expand, it is becoming more crucial to personalize treatment approaches to provide the most value to each individual patient. In addition, it is imperative to complement clinical trials with development of prognostic and predictive biomarkers. Prognostic biomarkers provide the likely natural course of MM in an untreated patient, while predictive biomarkers identify the subpopulation of patients who have better outcomes with specific treatment strategies. Over the last decade, researchers have explored new prognostic biomarkers, such as imaging (PET-CT) as well as assays that test for minimal residual disease (MRD), which is the small number of tumor cells that may remain after treatment. However, their utility as predictive biomarkers is not well established. The overarching aim of this proposal is to investigate risk- and response-adaptive clinical trial strategies for newly diagnosed MM while exploring the predictive role of these biomarkers.
We will apply this approach in three clinical trials for newly diagnosed MM. The first two trials compare various combinations of immunotherapeutic agents with current regimens. The first trial examines MRD assays and PET-CT as predictive biomarkers. The second trial assesses MRD to determine the need for transplant and maintenance. The third trial examines the effectiveness of the immunotherapeutic antibody Daratumumab in clearing the bone marrow of residual tumor cells before stem cell collection and transplant in MM patients having a less than ideal response to induction chemotherapy. Together, these studies may provide new therapeutic approaches and improve MRD assessment to better understand a patient’s response. These practice-changing strategies will help move the MM field towards personalized care.
Grant: 6541-18 | Translational Research Program (TRP):
Location:Baylor College of Medicine, Houston, Texas 77030
Project Title: Testing Targeted Therapy In Langerhans Cell HistiocytosisProject Summary:
Rationale and Background: Children with Langerhans cell histiocytosis (LCH) develop destructive lesions that can arise in virtually any organ including bone, brain, liver and bone marrow. LCH occurs with similar frequency as pediatric Hodgkin lymphoma, but there has historically been fewer opportunities for patients with LCH to participate in cancer research studies due to uncertain identity. LCH was first identified over 100 years ago, but only in the past ~5 years has been recognized as a disease in the family of pediatric cancers. Outcomes for children with LCH remain suboptimal, with over 50% failing to be cured with initial chemotherapy, and the majority of patients who are cured suffer long term consequences including problems with growth, control of hormones, and some develop a devastating progressive neurodegenerative condition. Patients who relapse or do not respond to front-line therapy are typically treated with highly toxic chemotherapy or eventually bone marrow transplant. Improved therapies are clearly needed for children with LCH.
Preliminary Studies: In 2010, a mutation in the BRAF gene (called BRAF-V600E), was discovered in over half of LCH tumor samples tested. Over the past 5 years, more mutations in the same cell growth pathway as BRAF (the MAPK pathway) have been discovered, accounting for over 85% of all cases of LCH. In the MAPK pathway, a series of proteins transmit messages from the cell surface to the nucleus of the cell, where that message is translated by turning on or off certain genes. In LCH, this MAPK pathway is overactive and never turns off, resulting in uncontrolled cell growth, resistance to cell death, and formation of destructive LCH lesions. Studies in blood cells from patients with LCH and in mice demonstrated that LCH is caused by activation of the MAPK pathway at specific stages of blood cell development. In early clinical trials with adult patients, LCH lesions responded to vemurafenib, a drug that blocks BRAF-V600E activation. Studies with cells from patients with LCH and experimental mice suggest that blocking MAPK pathway activation with drugs that inhibit MEK activation may be an effective therapeutic strategy for patients with LCH.
Hypothesis and Aims: We proposed to test the hypothesis that cobimetinib, which targets MEK activation, will be a safe and effective treatment for patients with refractory LCH, LCH-neurodegenerative disease, and disorders related to LCH that are also driven by MAPK activation. Additionally, we propose to study the responses associated with certain mutations, determine if cells in blood carrying LCH mutations can be used to follow disease activity, and study new mutations in patients who relapse despite cobimetinib therapy. This study will be carried out through a consortium of LCH disease experts at 11 different institutions through the North American Consortium for Histiocytosis Research.
Grant: 6552-18 | Translational Research Program (TRP):
Location:Walter & Eliza Hall Institute of Medical Research, Parkville 3050, Victoria
Project Title: Long-term In Vivo Imaging Of Bone Marrow Microenvironments In Multiple Myeloma.Project Summary:
White blood cells are soldiers of the immune system. These cells are responsible for surveillance of the body and protection from invading pathogens. When the machinery that controls growth and death of these cells is disrupted by genetic mutations, these cells can undergo massive unregulated expansion. This leads to the development of blood cancers such as leukemia and multiple myeloma (MM).
Blood cancer cells move uncontrolled throughout the body and expand to enormous numbers not normally present in healthy individuals. In addition, the cancer cells can secrete huge amounts of proteins that upset the equilibrium of healthy tissue in the body. In the case of MM, the leukemic cells infiltrate bones. This has dramatic consequences for the health of patients with MM. Cells that normally inhabit the bone are affected by overcrowding caused by expansion of cancer cells. This prevents them from performing their normal daily functions. For example, stem cells responsible for production of red blood cells that circulate throughout the body each day shut down and cannot make more cells leading to shortage of blood. MM cells can also damage the structure of the bones themselves leading to fractures and significant pain in over 80% of MM patients. Currently, this process is poorly understood. Unfortunately, there is no cure for MM and this is at least in part because MM can cells hide in the bone, protected from drugs used as treatment. Thus, considerable effort is needed to develop new treatments to overcome this resistance to treatment and manage the long-term effects of this disease on bone health.
We will solve this problem by watching how MM cells damage bone tissue using cutting edge microcopy. Using 3-dimensional printing technology, we produce custom optical windows that we surgically attach to living bone tissue. Through these optical windows we can view bone tissue for either short periods of time (hours) or throughout the entire disease process (weeks). Therefore, using this technology we will be able to see inside living organisms while MM cells grow, take over and then destroy bone tissue. Using our revolutionary approach, we are able to watch the same cells in the same bone tissue over hours and weeks. This will give us fundamental knowledge about the life cycle of MM and how it responds to treatment that have never been possible before. Once we can directly watch this process in action, we will be able to start to understand how MM cells live, destroy bone and evade therapy. Therefore, we will be able to develop new ways of targeting MM cells so that we can prevent bone damage, and even potentially stop the growth of MM cells leading to a cure.
Grant: 6548-18 | Translational Research Program (TRP):
Location:Dana-Farber Cancer Institute, Boston, Massachusetts 02215
Project Title: Development Of Histone Lysine Demethylase KDM3A Inhibitors For Multiple Myeloma TherapyProject Summary:
Cancer arises from a series of mutations in the DNA sequence that either activate (turn on) genes that allow cells to grow uncontrollably, or silence (turn off) genes that would normally tell a cell to die if it acquires DNA mutations. However, recent evidence suggests that some cancers inappropriately activate or silence genes through a different mechanism, called epigenetics. Epigenetics refers to chemical modifications to DNA and histone proteins that control gene activity without causing mutations in the DNA sequence. Recently, we found that one such epigenetic regulator, KDM3A, is overexpressed in multiple myeloma (MM). Biological investigation into the role of KDM3A in MM reveals that it directly regulates multiple other genes required for cancer cell survival, acting as a master regulator. Knockdown of the KDM3A protein in MM cells induces cell death and reduces tumor size in mouse models of MM. Likewise, knockdown of KDM3A reduces cancer cell interaction with the bone marrow, which is required for MM cell survival. Together, these findings suggest that KDM3A may be a novel therapeutic target for the treatment of MM, a disease that remains incurable.
Here, we propose to develop small molecule inhibitors of KDM3A in order to validate inhibition of KDM3A as a therapeutic opportunity in MM. We have developed an integrated chemical biology approach 1) to chemically synthesize and screen for novel KDM3A inhibitors, 2) to optimize inhibitors for drug activity and selectivity (on target effects) in cells and in animals, and 3) to validate KDM3A as a therapeutic target in cell and animal models of MM. Overall, the goal of this research is to develop small molecule inhibitors of KDM3A and to utilize them to gain a better understanding of how KDM3A drives MM biology, and to fully evaluate the therapeutic potential of KDM3A inhibition in MM. Thus, we have assembled scientists with multi-disciplinary expertise ranged from chemistry, medicinal chemistry, chemical biology and biology to achieve the goal proposed in this research. We envisioned that this research will provide the preclinical rationale to prompt clinical investigation of KDM3A inhibitors for MM which affects ~30,000 new patients a year. The identified small molecule inhibitors developed here will be further optimized for therapeutic use to improve patient outcome in MM.