Asking questions that lead to brilliant ideas is at the root of scientific discovery. Quantum leaps in science are made by this type of innovative thinking, but funding for the early pursuit of such initiatives is hard to obtain. However, the Pelotonia Research Award Program provides “idea” grants that allow creative teams of scientists to embark on research that could lead to discoveries resulting in better treatments and prevention strategies.
More than 32 research teams have received Pelotonia idea grants totaling millions of dollars. The teams represent collaborations among several colleges and departments, as well as three academic institutions (including Nationwide Children’s Hospital).
The awards are issued via a peer-review process conducted by scientists not competing for the grants. The grants cover an array of studies, from the genetics of triple-negative breast cancer to imaging of precancerous pancreatic lesions; from neurofibroma tumorigenesis and therapy to the molecular mechanisms of the body’s natural killer cells against multiple myeloma; from the role of the ATF3 gene in the development and treatment of chronic lymphocytic leukemia to genomic aberrations driving metastatic squamous cell carcinoma (a type of skin cancer).
2014 Pelotonia Idea Grants
Delivering an AML Drug in Nano-sized ‘Fat Bubbles’
Title: CD-33-Targeted Liposomal Bortezomib (aCD33-L-BZT) for AML Therapy
Principal Investigators: Robert Lee, PhD, and Andrienne Dorrance, PhD
Acute myelogeneous leukemia (AML) affects more than 14,500 Americans annually and has a poor survival rate. The drug bortezomib has potential to help AML patients, but it is only weakly effective against leukemia in its current form. In this project, an OSUCCC – James team from the colleges of engineering, medicine and pharmacy will develop a novel delivery system for this medication by packing the drug into nano-sized bubbles of fat and attaching it to a homing device that seeks out leukemia cells, sparing healthy cells. Preliminary studies suggest this approach effectively targets leukemia cells and results in lower drug toxicities. Data from the study will determine whether this approach is suitable for testing in humans.
Social Isolation’s Role in Breast Cancer Development and Progression
Title: Social Modulation of PTEN in Women
Principal Investigators: Courtney DeVries, PhD, Maryam Lustberg, MD, Cynthia Timmers, PhD
Studies show that women with breast cancer who are socially isolated have worse clinical outcomes. This OSUCCC – James team will examine whether loneliness and isolation alter cancer-related gene activity in breast tissue. The study investigates a molecular mechanism by which the social environment influences breast cancer initiation and progression. The team hypothesizes that a tumor-suppressor gene called PTEN plays a significant role in this process. Information from this study could reveal potential new diagnostic, therapeutic and prognostic tools for breast cancer prevention and treatment. Breast tissue for this study will be obtained from women undergoing biopsy at the Stefanie Spielman Comprehensive Breast Center for possible breast cancer.
Mental Health, Stress and the Response to Cancer Treatment
Title: Psychological and Inflammatory Responses in Relapsed and Refractory Patients with Chronic Lymphocytic Leukemia (CLL) Undergoing Ibrutinib Therapy
Principal Investigators: Amy Johnson, PhD, Barbara Andersen, PhD
Chronic lymphocytic leukemia (CLL) is the most prevalent form of adult leukemia and is currently incurable. This project will assess stress, depression and quality of life in patients receiving an effective new treatment called ibrutinib, which has been studied extensively in clinical trials at The OSUCCC – James. This study examines the relationship between cancer growth factors and patient psychological function. This information could help physicians make treatment decisions by identifying patients at risk for poor outcomes.
Biomarker-Based Two-Drug Therapy for Acute Myeloid Leukemia
Title: Phase I Study of AR-42 and Decitabine in Acute Myeloid Leukemia
Principal Investigator: Alison Walker, MD
Overall survival is low for both pediatric and adult patients with acute myeloid leukemia (AML) on standard chemotherapy. This study is a phase I (first-in-human) clinical trial to test a two-drug approach that could significantly increase remission in AML patients. Initial studies conducted at This OSUCCC – James, have shown that the drug decitabine is well tolerated in older AML patients and can achieve a 47 percent remission rate. Additionally, patients with higher levels of a substance in the blood called miR-29b had a better response to decitabine than those with lower levels. A second drug, known as AR-42, which was developed by OSUCCC – James researchers, increases levels of miR-29b in leukemia cells. This clinical trial will administer AR-42 first to AML patients as a way to increase miR-29b levels in the blood and possibly improve the effectiveness of decitabine therapy. The findings evaluate an innovative strategy for increasing the number of AML patients who achieve complete remission.
Studying Health Disparities in 100,000-Underserved in America
Title: Cancer Disparity Research Network (CDRN) Cohort Feasibility Study
Principal Investigators: Electra Paskett, PhD, Peter Shields, MD, Mira Katz, PhD, Paul Reiter, PhD, Eric Seiber, PhD, Mike Pennell, PhD
Despite an overall decrease in cancer incidence and death in many populations, significant health disparities exist in low income, racial and ethnic minority, rural, immigrant, under and uninsured and low-educated populations. This project will establish a cohort of 100,000 underserved people to better understand the causes of cancer disparities in the United States. The cohort will focus on four underserved population groups that studies have shown suffer from disparities: African Americans, Appalachians, Asians and Hispanics. This grant will support the formation of a coordinating center to collect and analyze data and biospecimens from The OSUCCC – James network of collaborating recruitment sites across the United States.
Targeting Oncogenes for New Liver Cancer Drugs
Title: Development of Novel Therapeutics Against Hepatocellular Cancer in Preclinical Models
Principal Investigator: Kalpana Ghoshal, PhD
Liver cancer is the third leading cause of cancer death in the United States and incidence rates are rising. The liver is designed to keep foreign substances out of the body, so developing drugs that effectively penetrate the liver and successfully target cancerous cells has been challenging. In this study, researchers will conduct preclinical tests to determine the effectiveness of new drugs that target two oncogenes—genes that promote cancer growth when highly expressed—along with a tumor-suppressing microRNA called miR-122, which is critical to maintaining normal liver function. Results from these studies could lead to a phase 1 clinical trial in liver cancer patients.
Understanding Molecular Crosstalk Driving Aggressive Breast Cancers
Title: Role of Slit in CXCR4-Mediated Breast Cancer Metastasis
Principal Investigator: Ramesh Ganju, PhD
Research suggests that two molecular pathways in particular play important roles in breast cancer development and how it is spreads, but little is known about the molecular conversations and the chain of events that lead to breast cancer growth and metastasis. A better understanding of this molecular crosstalk could help scientists identify points in the pathway to intervene and put the brakes on cancer development. This project seeks to further characterize the role of proteins in the two targeted pathways to better understand breast cancer growth, blood vessel formation and tumor spread. This information is especially critical for the development of new therapies in triple-negative breast cancers.
Brain Inflammation and Depression and Anxiety in Breast Cancer Patients
Title: Randomized Placebo Controlled Study of Minocycline for Amelioration of Chemotherapy Induced Affective Disorders (OSU 13165)
Principal Investigator: Courtney DeVries, PhD, and Maryam Lustberg, MD
Breast cancer survivors commonly experience depression and anxiety—particularly when undergoing chemotherapy. Inflammatory changes in the brain could be a primary cause of these symptoms. This OSUCCC – James team will study whether reducing inflammation in the brain using a readily available and well-tolerated drug called minocycline reduces depression and anxiety during chemotherapy. This study will be conducted in up to 30 postmenopausal women receiving chemotherapy for breast cancer at the Stefanie Spielman Comprehensive Breast Center.
Digital Image Analysis, Targeted Therapies for Glioblastomas
Title: Defining Molecular Events for Targeted Therapy in Glioblastoma Using Digital Image Analysis
Principal Investigators: Metin Gurcan, PhD, Jose Otero, MD, PhD, Brad Elder, MD, Vinay Puduvalli, MD, Jessica Winter, PhD
Glioblastomas are the most common and deadly of primary brain tumors. Despite aggressive treatment, glioblastoma patients live an average of 15 months. In this project, OSUCCC-James researchers are developing advanced image analysis techniques to help guide critical decisions in patient treatment before and after brain surgery. This technology could also guide personalized treatment options, based on the specific molecular characteristics of each patient’s tumor. Current imaging technologies make it difficult to distinguish between a cancer recurrence and treatment affected by chemotherapy and radiation. The goal of this study is to determine whether computerized image analysis combined with advanced protein analysis can significantly improve diagnostic accuracy and identify potential biomarkers that might help personalize treatment for each patient and provide insights into drug resistance.
Tackling Treatment-Resistant Prostate Cancer
Awardees: Qianben Wang, PhD; Steven Clinton, MD, PhD
When prostate cancer returns after surgery, it often no longer responds to drug treatment. For this study, OSUCCC – James researchers will use Pelotonia funds to identify genes that treatment-resistant prostate tumors need to grow and that could be potential new targets for prostate cancer drugs. The findings could lead to new treatments for prostate tumors that currently have no effective therapy.
Preparing for Resistance
Awardee: Sameek Roychowdhury, MD, PhD
Cancer happens in part because changes in certain genes cause cells to grow and divide when they shouldn’t. One of these genes is called the fibroblast growth factor receptor (FGFR). Researchers at the OSUCCC – James have designed a phase II clinical trial to test a new drug, called ponatinib, which inhibits hyperactive FGFR genes. However, cancer cells often develop resistance to the drugs that are used to treat them. Anticipating that resistance might develop in some patients during the ponatinib trial, the researchers have received a Pelotonia grant that will enable them to collect biopsy samples from each trial participant’s tumor before and after treatment. The grant will also enable the researchers to sequence 20,000 genes in each sample and look for new gene changes that could cause ponatinib resistance. The findings will provide a foundation for further research on how cancer cells become resistant to FGFR inhibitors and for the development of drugs to counter the resistant cells.
A Plant Component That Might Help Immune Cells Control Cancer
Awardee: Jianhua Yu, PhD
A Pelotonia grant is enabling an OSUCCC – James researcher to learn whether a substance from edible plants boosts the cancer-cell killing activity of a type of immune cell. The Pelotonia-funded study will investigate the ability of the plant substance, called phyllanthusmin C (PL-C), to stimulate the activity of natural killer (NK) cells. Ultimately, the researchers hope to show that PL-C in the diet will help NK cells control acute myeloid leukemia and perhaps other cancers.
Probing a New Target in Triple-Negative Breast Cancer
Awardees: Robert Brueggemeier, Harold Fisk, PhD; Chenglong Li, PhD; Pui-Kai Li, PhD; Yasuro Sugimoto
Triple-negative breast cancer (TNBC) is one of the most aggressive forms of breast cancer. It is defined by the absence of estrogen, progesterone and HER2 receptors. These molecules are targets for the drugs used to treat other forms of breast cancer. Without those targets, the usual breast cancer drugs are rendered ineffective, leaving no good treatments for TNBC. For this study, Pelotonia funding is enabling OSUCCC – James researchers to conduct laboratory studies to learn whether drugs that inhibit a molecule called Mps1/TTK are a promising treatment for TNBC and other aggressive forms of breast cancer.
A New Approach to Cervical-Cancer Prevention
Awardees: Paul Reiter, PhD, MPH; Mira Katz, PhD, MPH
Several types of human papillomavirus (HPV) cause cervical cancer and other types of cancer. Cervical cancer is largely preventable through regular screening, and current guidelines recommend that women ages 30-65 seek a Pap test and an HPV test every five years, or a Pap test every three years. But most women diagnosed with cervical cancer have had few or no Pap tests. One strategy for increasing the number of women screened for the virus is the use of HPV self-testing. Women collect samples by themselves at home and mail them in for testing. Self-testing might be particularly effective for screening women in underserved communities, such as Appalachia. For this study, OSUCCC – James researchers are using a Pelotonia grant to develop a pilot program for HPV self-testing among women in Appalachia who have undergone little if any cervical screening. The study will provide needed information about the value of self-testing for cervical cancer prevention.
Personalizing Multiple Myeloma Treatment
Awardees: Mitch Phelps, PhD; Ming Poi, Pharm D, PhD; Craig Hofmeister, MD; Susan Geyer, PhD
In 2012, 5,000 patients with multiple myeloma, an incurable cancer of the blood, were treated using stem-cell transplantation plus high doses of a drug called melphalan. The drug kills the person’s cancer cells, and the transplant rebuilds the person’s immune system. The treatment often prolongs patients’ lives and stops progression of their disease. But patients show dramatic differences in this progression-free period – from six months to 12 years. One problem is that individuals don’t metabolize melphalan the same way, resulting in differences in toxic side effects and differences in effectiveness from patient to patient. For this study, researchers are using Pelotonia funds to begin developing a step-by-step procedure, an algorithm, to personalize melphalan dosing to maximize the killing of myeloma cells while minimizing the drug’s harsh side effects
A Wearable Guidance System for Better Cancer Surgery
Awardees: Ronald Xu, PhD; Michael Tweedle, PhD; Alper Yilmaz, PhD
Tools that could help surgeons determine where a tumor ends and healthy tissue begins, and that could help detect hidden cancer cells, could greatly reduce cancer recurrence rates and improve the long-term outcomes of patients after cancer surgery. This Pelotonia-supported project is an initial step in developing such a tool. It will help develop and test a guidance system worn during surgery to identify surgical margins and guide the removal of tumors. The proposed system – the collaborative brainchild of clinical and engineering faculty – includes a fluorescence imaging module, surgical scene-capturing module, Google glass and a host computer. Prior to surgery, a cancer-targeting dye is injected into the patient’s vein. This dye is picked up on the fluorescence camera and fused with background images of the surgical area. Two additional cameras then reconstruct a 3-D topography of the surgical cavity and track the position of the surgical tool. Tumor margin and location information is further processed and projected to the Google glass, providing intraoperative imaging guidance.
A ‘Psychological Biomarker’ for Predicting Chemotherapy Side Effects
Awardees: Charles Shapiro, MD, and Kristin Carpenter
There is great variability in the side effects breast cancer patients experience with chemotherapy, and it remains difficult to predict a patient’s experience following chemotherapy. Clinicians believe that optimism (a general expectation of favorable outcomes) and coping (an individual’s reaction to perceived harm or threat) influence cancer patients’ quality of life, levels of fatigue, depression and sometimes even disease-free survival. But no clinical trial has yet evaluated whether optimism or coping in women with early-stage breast cancer can predict patients’ sense of chemotherapy-related side effects or of their health-related quality of life during or after treatment. This Pelotonia grant supports a clinical trial designed to learn if there is a correlation between chemotherapy treatment and side effects that include fatigue, nausea, vomiting, sensory neuropathy, pain, depression and insomnia. It will investigate whether mechanisms by which optimism and coping might influence these chemotherapy side effects. The findings could provide initial evidence of a “psychological biomarker” for predicting chemotherapy side effects, and they could assist in planning a larger phase III trial to test behavioral or other interventions that might lessen side effects via changes in optimism or coping.
Reversing Drug Resistance in Ovarian Cancer
Awardees: Jeffrey Parvin, MD, PhD, and David Cohn, MD
Chemotherapy kills cancer cells by damaging their DNA so badly that the cells cannot repair it. Nonetheless, ovarian cancer recurs in up to 80 percent of patients after treatment with chemotherapy. These OSUCCC – James researchers have found that a protein called histone deacetylase 10 (HDAC10) is part of an important DNA repair system in cells. They believe that this repair system allows some ovarian cancer cells to survive the damage inflicted by the platinum-based chemotherapy used to treat the disease. This Pelotonia grant will enable the investigators to examine whether drugs called HDAC inhibitors will knock out the HDAC10-powered DNA repair system and make drug-resistant ovarian cancers respond once more to platinum-based chemotherapy. If successful, the project will lay the groundwork for a new treatment strategy that might prolong the lives and reduce the suffering of women with ovarian cancer.
Arresting a Gene That Might Drive Esophageal Cancer
Awardees: Zui Pan, PhD; Tong Chen, MD, PhD
Esophageal cancer is the sixth leading cause of cancer death worldwide, largely because a large majority of cases are diagnosed at late stages of the disease. Research is needed to identify biomarkers for detecting the disease early and to develop new therapies for the disease. Studies by OSUCCC – James scientists have shown that a gene called Orai1, which helps regulate calcium levels cells, is present at abnormally high levels in esophageal cancer cells. The two researchers hypothesize that the hyperactive Orai1 gene causes abnormal changes in calcium levels in the cells and contributes to esophageal cancer progression and that inhibiting that over-activity could help control the disease. Their Pelotonia grant is enabling them to conduct experiments that will reveal more about the role of this gene in esophageal cancer and help them obtain larger grants for studies to learn whether drugs that target Orai1 would improve the treatment of esophageal cancer.
Targeting Two Genes Might Improve Melanoma Treatment
Awardees: William Carson III, MD; Albert de la Chapelle, MD, PhD; Kathrin-Ann Eisfeld, MD
Melanoma, the most deadly skin cancer, accounts for 75 percent of all skin cancer deaths in the United States, and its incidence is rising. Nearly 77,000 new melanoma cases were diagnosed in the nation in 2013. About 40 percent of all melanoma patients have a specific mutation in a gene called BRAF. The gene mutation also increases the likelihood that the cancer will spread to other parts of the body. Drugs have been developed that target mutated BRAF, but they work only in a subset of patients, most of whom ultimately become resistant to the drugs. This study uses Pelotonia funds to investigate BRAF-mutated melanoma and the role of a gene called microRNA-3151, a gene whose importance was discovered at the OSUCCC – James. The team’s initial data suggests that melanoma tumors that have mutated BRAF might respond better to BRAF inhibitors if microRNA-3151 is inhibited at the same time. This project will explore the mechanism of action of miR-3151 and evaluate whether BRAF-inhibiting drugs might more effectively treat melanoma when combined with inhibitors of miR-3151.
2013 Pelotonia Idea Grants
CD200R Signaling in Melanoma Progression and Immunotherapy
Awardees: Xue-Feng Bai and Lai-Chu Wu
Melanoma is the most dangerous type of skin cancer and the leading cause of death from skin diseases. Melanoma is characterized by early metastasis to distal organs, such as the lung, and the overall success of treatment in metastatic melanoma is limited. In our recent study we have found that melanoma cell expression of CD200, a cell surface glycoprotein, can significantly inhibit melanoma tumor formation and metastasis. Such inhibition of tumor formation and metastasis appears to be mediated by CD200 receptor (CD200R)-positive myeloid cells, since injection of B16 melanoma cells resulted in significantly accelerated tumor growth and metastasis in CD200R-deficient mice than in wild type mice. In addition, stimulation of CD200R in normal mice with an agonistic antibody in vivo dramatically inhibited metastatic B16 tumor foci formation in the lungs. Based on these observations, we hypothesize that CD200R signaling in myeloid cells inhibits melanoma tumor progression and that targeting CD200R is a useful immunotherapy of melanoma. To test this hypothesis, we will first generate CD200R-deficient Braf/Pten mice that develop conditionally inducible melanoma, to determine if lack of CD200R signaling in myeloid cells accelerates spontaneous melanoma formation and metastasis. Second, we will examine if triggering CD200R using an agonistic antibody is a useful approach for immunotherapy of spontaneous melanoma, and if so, determine what signaling pathway is involved. Information generated from these studies will lead to understanding the role of CD200R signaling in melanoma pathogenesis and immunotherapy.
CS-1 Targeted NK vs. T-cell Chimeric Antigen Receptor Therapy With or Without Elotuzumab
Awardees: Jianhua Yu and Craig Hofmeister, MD
Patients with relapsed hematologic malignancies represent an unmet therapeutic need. Chimeric antigen receptor (CAR) therapy represents an intriguing option for patients with multiple myeloma (MM) -- CS1 is a MM-associated antigen highly and universally expressed on MM cells but minimally expressed on normal cells, and thus can be utilized to direct cytotoxic immune cells to specifically kill MM cells. CAR NK cells are novel effector cells whose killing may be enhanced by monoclonal antibodies, and these cells may avoid the potentially lethal side effects of CAR T cells such as cytokine storms. The humanized CS1 mAb, Elotuzumab, can enhance antibody-dependent cellular cytotoxicity of NK cells against MM cells and has progressed to phase III trials. CS-1 targeted CAR NK cells are largely unexplored and their combination with a monoclonal antibody may be synergistic. We have generated CS1-CAR NK cells to more effectively eradicate MM cells in vitro. We hypothesize that targeting CS1 by CAR cells alone or in combination with CAR T cells or Elotuzumab is a promising therapeutic strategy, and the combination will be synergistic. We will test this by investigating the effect of CS1-CAR NK cells alone or in combination with CS1-CAR T cells or Elotuzumab in vitro and in an in vivo model system. We believe these experiments will lay the foundation for a phase I trial for relapsed myeloma patients utilizing autologous CAR NK cells.
Tethered Cationic Lipoplex Nanoparticle (TCLN) Assay for Early Lung and Liver Cancer Detection and Surveillance via Extracellular RNAs in Exosomes and Circulating Tumor Cells
Awardees: L. James Lee, Patrick Nana-Sinkam, Kalpana Ghoshal, Michael E. Paulaitis and Carl Schmidt
Lung cancer and liver cancer are two of the leading cause of cancer deaths worldwide. A patient-friendly early detection and surveillance method would substantially reduce the mortality in these serious diseases. ‘Liquid biopsy’ relying on detection of either circulating tumor cells (CTCs) or extracellular RNAs encapsulated in exosomes in patient blood samples has great potential to achieve this goal. But existing detection methods are far from perfection because CTCs are rare (~5-100 per 1 mL human blood) and exosomes are very small (<100 nm in diameter). Furthermore, none are able to simultaneously capture and detect both circulating exosomes and CTCs. We recently developed a simple and low-cost method ‘Tethered Cationic Lipoplex Nanoparticle (TCLN) biochip’ that, for the first time, may achieve this goal (a manuscript is under review in Nature Nanotechnology). In TCLN, molecular beacons (MBs) are pre-loaded in liposome nanoparticles to capture circulating exosomes and detect encapsulated extracellular RNAs from the patient blood or fluid sample without extra complicity. The in situ detection of target RNAs without diluting the sample leads to very high detection sensitivity not achievable by existing methods, e.g. qRT-PCR. The same biochip can also detect intracellular biomarkers in the captured CTCs by lipoplex internalization. Preliminary data in lung cancer cell line and patient blood samples using miR-21 and TTF-1 mRNA as biomarkers are very promising. TCLN can be extended to a multiplexing array design to allow the detection of many targets for RNA profiling. This new method may also be applied to other cancers and viral infection. In this proposal, we plan to evaluate its feasibility for early lung and liver cancer detection and surveillance. Our primary objectives are (1) to modify TCLN techniques so they can be used in flow cytometry and as a single-point detection method for clinical and point-of-care applications, (2) to compare target RNAs as biomarkers in tumor tissue and blood samples from MYC transgenic and miR-122 KO mouse models at different disease stages to evaluate the feasibility of using TCLN assay for early cancer detection, and (3) to identify target microRNA/mRNA biomarkers in lung and liver cancer and to conduct pilot studies using blood samples from well-defined lung and liver cancer patient groups. With data gathered from this project, we will submit two R01 or R33 proposals to NCI in two years and a P01 proposal in the future. This proposal is highly recommended by Nanoscale Science and Engineering Center (NSEC) at OSU. If funded, NSEC and the College of Engineering will match $50K/year for 2 years to allow the proposal team to pursue a broader scope of this challenging project.
STAT3 as a Mediator of Immune Suppression in the Pancreatic Cancer Stroma
Awardees: Greg Lesinski and Michael Ostrowski
One important hallmark of pancreatic cancer (PCa) is the dense stroma consisting of activated fibroblasts termed ‘pancreatic stellate cells’ (PSC) that surround each tumor. This fibrotic material arises due to chronic inflammation in the pancreatic microenvironment. Although these stromal cells produce numerous factors supporting tumor growth, little is known regarding their interactions with immune cells within the tumor microenvironment. Our data shows that PSC secrete factors that promote differentiation of immune cells into myeloid derived suppressor cells (MDSC). We hypothesize that STAT3 signal transduction in the tumor-associated stroma is essential for promoting immune suppression in PCa. Our long-term goal is to develop a comprehensive research program aimed at understanding how the stroma influences local and systemic immune suppression in PCa. This knowledge will be instrumental for prioritizing the most relevant stromal targets that could be manipulated in the clinic to enhance the efficacy of immunotherapy for PCa. To test our hypothesis we will 1) develop mice with spontaneously arising, mutant KRAS-driven pancreatic cancer [Mist1(KRasG12D/+)] and STAT3-deleted stromal fibroblasts and monitor tumor incidence and immune biomarkers and 2) define the external stimuli responsible for STAT3 activation in PSC, and identify other downstream pathways that cooperate with STAT3 to promote PSC survival. These data will be essential for a future R01 application.
Novel Small Molecule Inhibitor of PHD3 Effects on Human Breast Cancer Metastasis and Migration on Nanoscale Variable Modulus Devices
Awardees: Tim Eubank and John Lannutti
Better therapies for triple negative breast cancer patients are in critical need. Current options are surgery, taxane chemotherapies and radiotherapy. It is reported clinically that breast cancer patients whose tumors metastasize exhibit reduced desmoplakin (Dsp) expression. Dsp is the major protein component of the desmosome which regulates cell adhesion. Here, we introduce a novel small molecule inhibitor of PHD3, the prolyl hydroxylase that stabilizes HIF-2a, which we found augments Dsp mRNA and protein expression in PyMT and MDA-MB-231 tumor cells. Thus, we propose to study the ability of our novel PHD3 inhibitor to augment Dsp expression in human triple negative breast tumor cells implanted in SCID mice and reduce their migratory and metastatic ability. Key to this study will be the utilization of nanoscaled tools originating within the nationally known Nanoscale Science and Engineering Center (NSEC) at Ohio State. These tools will be used to quantify migration in response to chemotactic stimuli, differences in modulus representing age-based effects, gel-based barriers to cell migration to simulate invasion with and without embedded M2 macrophages, and DNA FRET-based ‘origami’ force sensors to potentially reveal local forces exerted by migrating tumor cells both in vitro and from tumor explants.
Insulin Receptor Splicing in Response to Hypoxia and Drug Resistance: A Pilot Study
Awardees: Dawn Chandler and Peter Houghton
Alternative pre-messenger RNA (pre-mRNA) splicing has emerged as a predominant mechanism for proteomic diversity and gene expression. This diversity has been emphasized by recent work that elucidates a network of alternatively spliced genes in response to cell stimuli that are part of the tumorigenic process (DNA damage and Epithelial to Mesenchymal Transition, EMT, for example). Though the importance of alternative splicing in a number of cell processes has been recognized since its discovery, the mechanisms underlying its regulation remain poorly understood. Solid tumors are characterized by both transient and chronic hypoxia, and the ability for tumor cells to adapt to hypoxia is essential for tumor progression. We have shown recently that stimulation of vascular endothelial cell proliferation and angiogenesis induced by the major angiogenic factor, VEGF, is dependent upon insulin-like growth factor (IGF) signaling. Additionally, both human vascular endothelial cells and cells derived from pediatric sarcomas express predominantly the alternatively spliced form of the Insulin Receptor gene (IN-R), that has high affinity for IGF-2. Importantly, IN-R is spliced in response to hypoxia, and pediatric sarcoma cells secrete IGF-2. Consequently, IN-R splicing appears critical in progression of pediatric sarcomas both from acting as a receptor for autocrine growth of tumor cells, and for paracrine growth of vascular cells, and hence angiogenesis. The primary goal of this proposal is to understand the mechanisms and consequences of IN-R pre-mRNA splicing in response to hypoxia. This work will test the hypothesis that regulatory elements and splicing factors are modulated in response to hypoxia and are involved in the alternative splicing of IN-R to contribute to tumorigenesis. In order to determine the effectors of IN-R hypoxia-induced splicing, we have developed a novel in vitro splicing assay that models pre-mRNA splicing from cells that have undergone hypoxia. We will utilize the hypoxia-induced splicing assay to identify RNA sequences and their respective binding partners that are necessary for regulation of IN-R pre-mRNA splicing. Furthermore, we will use established mouse xenograft assays and novel antisense oligonucleotides (ASOs) to modulate IN-R splicing and determine the role of the spliced isoforms in tumor progression and as novel targets for therapeutic intervention.
PRMT5 Dysregulation as a Driver Event in Richter’s Transformation
Awardees: Rosa Lapalombella and Robert Baiocchi
Richter's transformation (RS is a complication of B cell chronic lymphocytic leukemia (CLL) in which the leukemia changes into a rapidly proliferating diffuse large B cell lymphoma (DLBCL)1 with a poor prognosis despite the use of standard lymphoma therapy. The mechanisms underlying this transformation process remain to be clarified. PRMT5 is a type II methyltransferase that is highly expressed in lymphoma cell lines and primary tumors but not in normal resting or proliferating B cells. PRMT5 has been shown to induce transcriptional repression of cell cycle regulatory and tumor suppressor genes and is considered a driver event in both initiation and maintenance of cellular oncogenesis. Our preliminary data show that PRMT5 is variably expressed in CLL and overexpressed in patients with RS months prior to the morphologic transformation process where large B-cell lymphoma is noted. We hypothesize PRMT5 dysregulation to be both a driver event in CLL transformation to DLBCL and a potential therapeutic target for intervention. By using ChiP-Seq and RNA-Seq methods, we aim to provide insights of how PRMT5 dysregulation drive key cell growth and survival programs required for initiation and maintenance of the CLL and RS phenotype. This information will be vital to develop novel therapeutic approaches for RS, a currently incurable disease.
Gene Discovery Using a Drosophila Tumor Model
Awardees: Amanda Simcox and Vicor Jin
The Ras signaling pathway is implicated in multiple cancers, correspondingly, there is an intense effort to identify Ras regulators and effectors. We have developed a conditional Ras-driven tumor cell model in Drosophila. When oncogenic RasV12 is expressed the cells proliferate, and when RasV12 is switched off the cells become dormant, but remain viable for weeks. Our goal is to use this model to discover new genes in the Ras pathway and to investigate tumor dormancy. RNAseq data comparing cells in the two states, RasON and RasOFF, has identified 363 conserved genes that are significantly induced by Ras expression and 300 conserved genes that are specifically expressed in dormant cells. Functional analysis of a subset of prioritized genes from each set will be conducted using RNAi. In addition to finding new genes in the Ras pathway, it is important to understand tumor cell dormancy because in this quiescent state the cells evade cancer therapies targeting proliferating cells. These dormant tumor cells, however, need to be killed too because later they can cause recurrence of the cancer.
The Role of the Epstein Barr Virus in NK-cell Lymphoma
Awardees: Aharon G. Freud, Robert Baiocchi and Pierluigi Porcu
Extranodal NK-cell lymphoma, nasal type (ENKL) is an aggressive, uniformly EBV-positive extranodal non Hodgkin lymphoma (NHL) of natural killer (NK) cell origin that affects immune competent individuals in their 40-SOs. Even patients with disease localized to the nasopharynx typically experience early dissemination to other extranodal sites, despite aggressive chemo-radiation. The disease is rare in the US but is common in Latin America. While the mechanism of EBV-induced B-cell transformation in immunosuppressed patients has been well studied, very little is known about how EBV infects and transforms NK cells, what subsets of NK cells are targeted, what EBV genes are expressed in tumor cells, what are the EBV proteins recognized by the host T-cells, and how the latter affect outcome. Even less is known about the mechanisms of drug resistance that so greatly limit the efficacy of systemic chemotherapy in ENKL. Therefore, models to study viral pathogenesis, immune response, and drug therapy are needed. We hypothesize that EBV-positive NK lymphoma cells (EP-NK) circulate in the peripheral blood (PB) of ENKL patients and that these cells can be identified and purified to aid in the development of investigational models of ENKL. We also hypothesize that ENKL tumor and circulating lymphoma cells express EBV antigens that can be recognized by the host's T-cells. Our key objectives therefore are to develop models to study mechanisms of EBV-induced NK cell transformation in vitro and in vivo, and to characterize the host's immune response to ENKL for therapeutic purposes. Our group has a demonstrable collective expertise in the clinical and laboratory aspects of EBV-associated lymphomas, including ENKL, and in the biology of normal NK cells. The clinical resources to complete the proposed aims are available. Multiple vials of cryopreserved PB mononuclear cells (PBMC) from 8 OSU ENKL patients obtained at the time of detectable EBV viremia are available in the OSU LTB for specific aim 1 (purification of EP-NK cells) and specific aim 2 (study ofT-cell response). We have also partnered with investigators from the Institute Nacional de Enfermedades Neoplasicas (INEN, Peru') and with Dr. Natkunam (Stanford), who is internationally known for her work in ENKL and has an ongoing research partnership with colleagues in Guatemala. INEN will provide tissue blocks or slides and Dr. Natkunam will provide a tissue microarray (TMA) with >100 cases of ENKL for specific aim 2 (study of EBV gene expression in ENKL tissue). Genomic DNA from these samples will also be obtained for later experiments, which include next generation sequencing (NSG) of viral and tumor cell DNA and RNA and methylation profiling.