Grant: 5463-18 | Career Development Program (CDP):
Location:La Jolla Institute for Allergy and Immunology, La Jolla, California 92037
Project Title: TET Proteins In B Cell Function And Malignancy.Project Summary:
DNA methylation is a fundamental biological process that controls the activation state of genes, which represent basic modules of biological systems. TET proteins were recently identified as enzymes that mediate the removal of DNA methylation and have been shown to play vital roles in normal development of organisms. Moreover, the activity of TET enzymes is often deregulated during the pathogenesis of various hematological and non-hematological cancers. Consistent with these observations, recent studies have identified loss of TET protein functions in B-cell-derived malignancies, which are the most common hematological malignancies among adults in Western countries. However, the precise role of TET proteins in the regulation of normal B-cell functions and the mechanisms by which loss of TET activity contributes to development of B-cell-derived malignancies remains unclear. Interestingly, my preliminary studies suggest that TET protein shave important roles in the normal functions of B-cells and that loss of TET expression and activity in B-cells is associated with increased instability of the genetic material, which may cause a predisposition to cancer. I am currently investigating the mechanisms by which TET proteins control the physiological functions of B-cells as important components of the immune system. I will further extend these findings to understand mechanisms by which loss of TET proteins contributes to the development of B-cell lymphoma. Because TET protein functions are frequently deregulated in various cancers, the insights gained from these studies may be important in deciphering the origins of not just B-cell malignancies but also other kinds of cancers associated with deregulation of TET protein functions. This knowledge can be further applied in future designing of specific therapies that may potentially be efficacious across a broad spectrum of hematological and non-hematological malignancies.
Grant: 3376-18 | Career Development Program (CDP):
Location:The University of Utah, Salt Lake City, Utah 84112-9003
Project Title: Determining The Role Of SIRT5 In Acute Myeloid LeukemiaProject Summary:
Acute myeloid leukemia (AML) is the most deadly blood cancer, with more than 70% of patients dying from the disease within five years after diagnosis. The treatment option shave remained largely unchanged for the past 30 years. Chemotherapy and stem cell transplant are still the standard therapy for AML. The fact that most patients with AML will eventually relapse and succumb to their disease defines an urgent, unmet medical need for more effective drugs to treat this disease. To answer this call, we have taken a novel approach to identify new drug targets. We have methodically evaluated 1200 cancer-related genes one by one in patient samples to determine which genes are vital to the survival of AML cells. We have found that in approximately 70% of patient samples and cell lines, removal of SIRT5, a gene that regulates energy metabolism, kills AML cells but not healthy cells, suggesting that SIRT5 may be a new drug target in AML. The overall goal of my research is to better understand the role of SIRT5 in AML. Using a larger cohort of primary AML cells donated from patients, I will determine how common SIRT5 dependence is in this larger group of patients. My preliminary data also suggests that the dependence of AML cells on SIRT5 is not simply a matter of increased expression of the gene. Therefore, I will perform experiments to determine what other genes are involved in SIRT5-mediated dependence of AML cells. I will also determine how SIRT5 promotes a pro-tumor energy metabolism in AML cells and how inhibition of this abnormal metabolism kills AML cells. The aims of my proposal are to answer the following questions: 1) How common is SIRT5 dependence in AML, and what is the genetic basis of this dependence? 2) How does inhibition of SIRT5 kill AML cells? My proposal has the potential to establish SIRT5 as a novel drug target in AML and lead to the development of a new class of drugs that target SIRT5 to improve patient outcomes in AML.
Grant: 6529-18 | Translational Research Program (TRP):
Location:University of Minnesota, Twin Cities, Minneapolis, Minnesota 55414
Project Title: Innate Lymphoid Cell Type 2 Infusion For Graft-versus-host Disease (GVHD) Prevention And TreatmentProject Summary:
Hematopoietic stem cell transplantation (HSCT) using donor cells transplanted into patients can potentially cure patients with hematological malignancies. However, an immune response of the donor cells against the patient, termed graft-versus-host disease (GVHD) can lead to death in the patient limiting the use of stem cell transplantation. Here, we have discovered that a unique population of cells, known as innate lymphoid type 2 (ILC2s) have a potent capacity to prevent and treat murine acute GVHD. ILC2s are located at mucosal surface sites of exposure to foreign antigens and pathogens, especially the gut and the lung, which are key GVHD target organs. ILC2s control adverse immunological events by releasing anti-inflammatory cytokines and tissue reparative factors, as well as activate a network of immune regulatory cells in the recipient. We have found that conditioning regimens used to prepare patients for HSCT eliminates these cells before transplant and that such elimination predisposes the recipient to GVHD. Indeed, in HSCT patients, the slow tempo of recovery of ILC2s correlates with an increased likelihood of conditioning regimen toxicity and GVHD. In a mouse GVHD model, we have shown that interleukin-33, which increases ILC2s, when given prior to conditioning can markedly augment ILC2s and reduce GVHD. Further ILC2s given at the time of HSCT migrate to the gut, a primal GVHD target organ, and reduce GVHD. In mice with active acute GVHD, ILC2s given in 2 doses were remarkably effective in ameliorating lethality. Moreover, ILC2 infusion did not impair the graft-versus-tumor response . Thus the net result was control of GVHD without adverse effects on anti-leukemia responses. Using umbilical cord blood or cytokine mobilized peripheral blood, stem cell sources that have been used to replace the need for bone marrow harvests, we have developed a clinical relevant process to isolate and expand human ILC2s with validated ILC2 cell surface and functional characteristics. The present proposal will optimize the production of ILC2s from these 2 sources, ensure the process is FDA compatible, choose the preferred ILC2s source, and test the preferred product to suppress human peripheral blood cell mediated GVHD in vivo with mice that have been reconstituted with a human immune system. The project will then move to an FDA approved facility at one of the 2 sites (University of Minnesota for cord blood or University of North Carolina-Chapel Hill for peripheral blood) to perform the large scale validation runs required for FDA approval of a clinical trial. The competitive renewal will proceed with a combined two institution clinical trial of ILC2 infusion to treat patients with active acute GVHD that have failed to respond to standard of care as these patients have a very poor prognosis.
Grant: 6549-18 | Translational Research Program (TRP):
Location:The University of Iowa, Iowa City, Iowa 52242
Project Title: Clinical Investigation Of CD38+/ Light Chain+/CD24+ As Putative Multiple Myeloma Stem Cell MarkersProject Summary:
Treatment failure in cancers, including, multiple myeloma (MM), is due to persistence of a minor population of cancer stem cells (CSCs), which are mainly non-cycling and very drug-resistant tumor cells. One major clinical observation supporting the existence of MMSCs is that we have shown that gene expression profiles (GEP) remain abnormal in many MM patients with long-lasting complete remission (CR) (> 10 years), suggesting the persistence of a cancer cell population with very low proliferative capacity and very limited sensitivity to our most intensive therapies. Different groups have reported on the presumed identity of MMSCs; however, a unique MMSC phenotype has not yet been established. To further identify phenotypic markers of MMSC, gene expression profiles were analyzed using primary MM cells of both MM stem cells and the bulk MM cells. The cell surface protein CD24 was found to be significantly upregulated in the MMSCs compared to the bulk MM cells. We have confirmed that CD24+ MM cells showed stem cell features, such as increased clonogenic potential, drug resistance and tumor formation capacity with as little as 10 cells. Importantly, we also confirmed that CD24+ MM cells exist in clinical MM samples, which are CD38+/CD24+/k+ or λ+ . Therefore, we have designed two Specific Aims to accomplish this project: (1) We will characterize the role of CD24 in maintaining stem cell features and investigate its clinical relevance in MM; and (2) We will determine whether CD38+/CD24+/k+ or λ+ primary MM cells have cancer stem cell features. Improved understanding of MMSCs biology will likely lead to the development of novel therapeutic targets that can be tested in the laboratory and in the clinic. Evidence from clinical trials and correlative studies should provide proof that that inhibition of MMSCs leads to improvement in long-term clinical outcomes. Such critical studies can only be conducted if we are able to determine and analyze if CD38+/CD24+/k+ or λ+ are indeed consistent MMSC markers. Eventually, we hope this work can define MMSC markers in clinical samples and to use this knowledge to develop a novel therapy to prevent MM relapse.
Grant: 6542-18 | Translational Research Program (TRP):
Location:Oregon Health & Science University, Portland, Oregon 97239
Project Title: Novel Approach To Thwart MYC In B-cell Neoplasia By Selective Targeting Cyclin-dependent Kinase 9Project Summary:
Diffuse large B-cell lymphoma (DLBCL), a disease of the lymph nodes, is the most common subtype of non-Hodgkin lymphoma accounting for >10,000 deaths in the United States annually. While “targeted therapy” has made significant progress in treatment of blood cancers, we continue to treat DLBCL with standard chemotherapy regimens, which are associated with significant side effects and high rate of failure. When DLBCL recurs after initial therapy, it often becomes incurable with chemotherapy. The novel class of agents called inhibitors of cyclin-dependent kinases (or “CDK inhibitors”) has shown promise in therapy of cancer. Cyclin-dependent kinases are proteins which exist in cells under normal conditions. A whole range of them exist and ensure that cells can make all the necessary components for their survival, as well as reproduce. Cancer cells co-opt CDKs in their unlimited growth, and thus CDKs are attractive targets in cancer therapy. However, drugs which inhibit multiple CDKs are toxic, probably because they inhibit he function of multiple CDKs in normal (non-cancerous) cells. By contrast, emergent selective CDKs which target only one-two specific proteins hold promise to be more efficacious and have fewer adverse events. Here we propose to study how selective CDK inhibitors work in cancer, specifically focusing on inhibition of CDK9. Our preliminary experiments suggest that selective CDK9 inhibition stems lymphoma growth by disrupting the function of another protein, called MYC. MYC regulates synthesis of many cellular constituents which ultimately ensure tumor survival and growth. MYC levels are high in DLBCL and it contributes to therapy resistance. Here we will study the effect of CDK inhibitors on MYC function. We will determine how MYC function is affected, and whether MYC disruption is the dominant mechanism of how cells die in response to CDK inhibitors. We will also search for drug partners which will make CDK inhibitors more toxic specifically to tumor cells. Together, this study will help develop novel targeted therapies with the goal of eradicating DLBCL.
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: 5473-18 | Career Development Program (CDP):
Location:Broad Institute, Inc., Cambridge, Massachusetts 02142
Project Title: Probing The Translatome For Neoantigens In Chronic Lymphocytic LeukemiaProject Summary:
Our cells display on their surface a snapshot of their protein repertoire processed and presented in the form of antigens. Antigens are recognized by the immune cells, which assess whether the antigens are known or foreign and mount an immune response against cells with foreign antigens. DNA mutations in cancer cells produce mutant proteins, presented as neoantigens on the cancer cell surface. The immune cells recognize neoantigens as foreign and attempt to mount an immune response against these cancer cells. Recently approved immunotherapy drugs boost this immune response and show great promise in the clinic, but the response is often non-specific and may affect healthy tissues, resulting in autoimmunity. Thus, directing the immune cells to defend against specific neoantigens will further boost the immune response and minimize autoimmunity. However, identifying and selecting neoantigens to target remains challenging.
Putative neoantigens are commonly predicted by sequencing and identifying mutations in the exons (molecules which mostly contain information coding for amino acids) of a given genome. This approach is generally sufficient for cancers characterized by many DNA mutations. However, some cancers, such as chronic lymphocytic leukemia (CLL) accumulate fewer DNA mutations, resulting in fewer predicted neoantigens. Since exons represent about 1% of our genome and mutations can be found anywhere in the genome, we have the opportunity to identify previously unknown neoantigens that are generated from mutations outside of the exons. Using next generation sequencing technologies and computational tools, we will expand the neoantigen search beyond currently known genes, maximizing our chances at identifying neoantigens to target. The expanded search for neoantigens will broaden potential targets for therapy for CLL. Importantly, the computational approaches we are developing may be useful beyond CLL and may be used to identify novel neoantigens in any cancer, particularly those with low mutation load.
Grant: 5467-18 | Career Development Program (CDP):
Location:Walter & Eliza Hall Institute of Medical Research, Parkville 3050, Victoria
Project Title: The Key To Cancer Cell Death; Regulation Of The Pro-apoptotic Protein BIMProject Summary:
Treating blood cancer patients with conventional approaches remains unsatisfactory because the cancer often recurs and because of the undesirable side effects caused by many of the treatments currently offered. By improving our understanding of the genetic basis of hematopoietic malignancies, we can develop targeted agents that are more selective in their action against tumor cells.
In healthy organisms, many cells have a finite lifespan. These cells undergo a normal, programmed cell death process called “apoptosis” and are then replaced with new cells. Apoptosis is regulated by a number of proteins, including pro-apoptotic proteins, such as BIM and anti-apoptotic proteins, such as BCL2. A prominent hallmark of many leukemias and lymphomas is their inability to undergo apoptosis, allowing them to survive indefinitely. Research at my host institution contributed to the development of venetoclax, an inhibitor of the pro-survival protein BCL2, which treats cancers by restoring their ability to undergo cell death. In chronic lymphocytic leukemia (CLL), a leukemia where BCL2 is overactive, remarkable results have been achieved in clinical trials, recently leading to FDA approval of the drug for treatment of patients with high-risk CLL. Since different tumor types may express different apoptotic regulators, the results with venetoclax suggest that targeting apoptotic regulators specific to a tumor type may lead to better patient outcomes in a number of different cancers.
In mantle cell lymphoma (MCL), a disease related to CLL, expression of the pro-death molecule BIM is suppressed in 20% of patients. Thus, restoring BIM expression should sensitize these cells to undergo cell death, which may be particularly beneficial in combination with standard-of-care chemotherapeutic agents or the BCL2 inhibitor. This combination therapy may result in greater cell death and thus an enhanced therapeutic benefit. Though the role of BIM in regulating cell death is well-described, it is less clear how levels of BIM are regulated in normal cells or how BIM is suppressed in MCL. The focus of my research is to unravel the molecular mechanisms by which the expression of pro-death molecules, such as BIM, is controlled. I anticipate that the results I obtain could help explain how hematopoietic cancer cells can evade cell death and potentially lead to the development of novel approaches to treat these malignancies more effectively.
Grant: 3373-18 | Career Development Program (CDP):
Location:Emory University, Atlanta, Georgia 30322-4250
Project Title: Tetrameric Acetyltransferase ACAT1 Is A Novel Therapeutic Target In Treatment Of Human LeukemiaProject Summary:
Tyrosine kinases (TKs) are a group of proteins that serve as "on/off" switches to control various cellular functions. When TKs are continuously turned on (“activated”) in blood cells leukemia may result. Anti-leukemia drugs that are designed to target TKs in leukemia cells are widely used in clinics. However, many patients develop resistance to these drugs over time, making them less effective or not effective at all. Thus, it is critical to find alternative therapies to battle drug resistance and improve clinical outcome. Cancer cells, including leukemia cells, consume more sugar compared to normal cells. However, unlike normal cells that use glucose primarily for energy, cancer cells use glucose primarily to generate the “building blocks” of proteins and lipids, enabling the cancer cells’ rapid division rate. A protein complex called pyruvate dehydrogenase complex (PDC) functions as a “gatekeeper” to control the flow of sugar toward either energy generation or building block production. When the PDC is “closed,” such as in leukemia cells, most glucose will go towards the production of building blocks. Thus, finding out how PDC activity is inhibited in leukemia cells may uncover a new vulnerability of human leukemias.
Our previous research showed that a protein called acetyl-CoA acetyltransferase 1 (ACAT1) can significantly inhibit PDC activity. Our current studies show that ACAT1 can be highly activated in leukemia cells by TKs through a chemical modification called tyrosine phosphorylation, resulting in a more stabilized and activated form of ACAT1. Most importantly, we found that ACAT1 is more activated in leukemia patients than in healthy people, resulting in further PDC inhibition. Thus, we think ACAT1 is a promising anti-leukemia drug target. By screening FDA-approved compounds, we successfully identified a novel ACAT1 inhibitor called Arecoline Hydrobromide (AH). Our results showed that AH can effectively inhibit leukemia cell growth. We will perform further studies to gain a detailed understanding of how AH inhibits PDC, and we will test AH potency in various leukemia cell lines, animal models, and leukemia patient samples. We will also work with expert chemists to develop new ACAT1 inhibitors, related to AH, but with improved potency. Altogether, this research will provide new insight into anti-leukemia drug design, help identify new novel drug targets, and ultimately uncover novel therapeutic approaches in blood cancers.
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.