Katie M. Reindl, Ph.D. - US grants

Biological Sciences (61). The Virtual Cell development team is expanding its existing collection of high-quality, animation-based educational modules that feature molecular and cellular processes (http://vcell.ndsu.edu/animations). These animations are freely accessible from the VCell download site (http://vcell.ndsu.nodak.edu/animations/downloads/), and are widely used by teachers as stand-alone videos or incorporated into personal learning tools. The publicly available NDSU Virtual Cell Channel on YouTube is used by learners at all levels (http://www.youtube.com/user/ndsuvirtualcell) to better understand these cellular and molecular processes. This project is expanding the VCell collection by 1) developing additional animations for widespread (including international) distribution; 2) creating Biology-in-the-Hand applications, pioneering WWW-delivered learning modules for use with portable media players; 3) broadening the utility of the animations by offering the narration in other languages; and 4) utilizing a recently developed National Educational Media research framework to better understand those aspects of animations that aid learning. In accomplishing these goals, the sponsoring institution is collaborating with three partner institutions that have previously developed or used animations for biology education: East Carolina University, the University of Colorado, Denver, and the University of Toronto. These institutions are also serving as tests sites to measure the impact of specific animation designs on student learning. The major product from this project is a suite of high quality animations that can be accessed as stand-alone learning tools or embedded in learning applications that can be downloaded and accessed anytime/anywhere by a diversity of teachers and learners. Development of the animations is being guided by learning research that provides for better understanding of those components of an animation that aid student learning and that allow for effective delivery on mobile devices. The multiple-design concepts being incorporated by the project are making the animations attractive to a larger learner base, and allowing teachers to reconsider how complex processes may be taught.

? DESCRIPTION (provided by applicant): Although over half of all lung cancer patients receive radiation therapy as part of their treatment plan, the therapeutic window of radiation is quite small, and it is difficult to deliver tumoricidal doses of radiation without the development of severe adverse effects in the nearby normal tissues. An agent that could protect healthy lung tissue from radiation toxicity and at the same time sensitize lung cancer tissue to radiation therapy, would be a major advance in the treatment of lung cancer, the leading cause of cancer deaths in the United States. The long term goal for this research is to develop clinically-useful, natural product-based agents that improve treatment for cancer. The objective for this application is to establish the dual action of a natural product as both a tumor cell radiosensitizr and a normal cell radioprotector by determining its mechanisms of action and therapeutic efficacy in lung cancer radiotherapy. The central hypothesis is that the natural product PPLGM sensitizes tumor cells to radiation-induced cell death through inhibition of HIF2a and induction of ROS, and protects healthy cells from radiation by enhancing their antioxidant response. To test the central hypothesis, two Specific Aims are proposed: 1) Establish the mechanisms for PPLGM-induced lung cancer cell radiosensitization and healthy lung cell radioprotection in vitro; and 2) Evaluate the therapeutic efficacy of PPLGM for lung cancer radiosensitization and healthy lung cell radioprotection in vivo. For the first aim, functional assays will be performed t identify the lung cancer cell genotypes for which PPLGM is a radiosensitizer and the healthy lung cell types for which PPLGM is a radioprotector. Mechanistic studies will be done to evaluate the involvement of HIF2a in PPLGM-induced ROS, DNA damage, and cell death for lung cancer cells, as well as transcriptional targets of the hypoxic and antioxidant responses in lung cancer and healthy lung cells. For the second aim, the radiosensitizing effects of PPLGM on lung tumors will be established in a mouse model. Additionally, the radioprotective role of PPLGM will be evaluated in a mouse model of thoracic radiation-induced pneumonopathy both short-term (1 month) and long-term (4 months). The approach is innovative because it uses a novel dietary agent that shows potent anti-cancer effects with protective effects in healthy tissues for the first time in the context of lung radiotherapy. The proposed research is significan because it is expected to advance the field of radiation therapy by development of a more effective treatment for lung cancer. The results from this project are also expected to expand understanding of how natural products act as radiosensitizers for cancer cells and, at the same time, radioprotectors for healthy cells. Such knowledge has the potential to change the field of radiation therapy and result in more effective treatment for many cancers.

__________________________________________________________________________________________________ Project-3. Combination Therapy: Targeting Pancreatic Cancer with a ROS Inducer and Gemcitabine (PI: Dr. Katie Reindl) Project Summary Pancreatic Cancer (PC) is an extremely deadly disease with a mortality rate of nearly 95%. The current treatment options available for PC patients only extend their lives by a few months. Therefore, there is a critical need to identify potent compounds that could enhance the effectiveness of chemotherapies, especially for K- ras mutant PC cells that are often resistant to treatment. The goal of this research project is to evaluate a novel natural product-based agent in combination with gemcitabine (GEM) for pancreatic cancer therapy, eventually leading to the ultimate development of clinically-useful natural product-based agents for the treatment of cancer. The project objective is to establish piperlongumine (PPLGM) as a chemosensitizer for PC by determining its mechanisms of action and therapeutic efficacy. Our central hypothesis is that PPLGM, isolated from the fruits of long peppers, sensitizes tumor cells, but not normal cells, to chemotherapy-induced cell death through induction of reactive oxygen species (ROS) and enhanced DNA damage. We have formulated this hypothesis based on the existing literature and our own preliminary findings that show PPLGM elevates ROS levels in cancer cells leading to enhanced tumor cell death. To test our central hypothesis, we propose three Specific Aims: 1) To investigate the mechanisms by which PPLGM enhances ROS levels and induces cell death in PC cells; 2) To evaluate the effect of PPLGM on sensitizing PC cells, but not normal cells, to chemotherapy; and 3) To evaluate the therapeutic efficacy of PPLGM alone or in combination with GEM in mouse models of PC. For the first aim, we will treat normal and PC cells that are K-ras mutant and wildtype with PPLGM, and we will determine the molecular mechanisms by which PPLGM causes ROS-induced cell death. For the second aim, we will use the same cells to determine the combined effect of PPLGM and GEM on PC cell death and investigate their synergistic mechanisms for inducing PC cell death. For the third aim, we will evaluate the therapeutic efficacy of PPLGM alone or in combination with GEM using an orthotopic mouse model of K-ras mutant human PC and a transgenic mouse model. The approach uses a novel dietary agent that shows potent anti-cancer effects in the context of chemotherapy for PC. The proposed research is expected to vertically advance and expand understanding of how a ROS-inducing agent can be used as a chemosensitizer for cancer treatment. Ultimately such knowledge has the potential to change the field of chemotherapy and result in more effective treatment for many cancers. __________________________________________________________________________________________________

PROJECT SUMMARY Less than 9% of pancreatic ductal adenocarcinoma (PDAC) patients survive 5 years beyond diagnosis. A primary reason for the low survival rate is that the best available treatments are not that effective for many patients, and many tumors become resistant to them. A treatment approach that could sensitize tumor cells to chemotherapy would be a major advance in the treatment of PDAC, the most deadly of the major cancer types in the United States. The long term goals for this research are to enhance the biomedical research environment at North Dakota State University and to identify clinically-relevant therapeutic targets to improve treatment outcomes for PDAC patients. The objectives for this application are to provide training opportunities for undergraduate and graduate students and to establish the tumor-promoting roles of the antioxidant enzyme glutathione S-transferase pi 1 (GSTP1) in PDAC models. The central hypothesis is that inhibition of GSTP1 can sensitize PDAC cells to platinum-based chemotherapy and alter gene and metabolite expression profiles that limit tumor growth. To test the central hypothesis, three Specific Aims are proposed: 1) Determine the chemosensitizing effects of GSTP1 inhibition for PDAC cells, 2) Identify the gene expression changes associated with GSTP1 inhibition in PDAC cells, and 3) Identify the metabolome changes associated with GSTP1 inhibition in PDAC cells. For the first aim, cell culture and mouse models will be used to determine the degree to which pharmacological and functional GSTP1 inhibition influence sensitivity to platinum-based chemotherapy. For the second aim, our transcriptome data will be validated in GSTP1 knockdown PDAC cell lines and in PDAC cells treated with a GSTP1 inhibitor. Functional assays will be performed to further identify anti-tumor mechanisms associated with GSTP1 inhibition. For the third aim, targeted metabolomics will be used to determine the metabolite expression changes in PDAC cells when GSTP1 function is impaired. The approach is innovative because it will involve evaluating a molecular target that has not been studied before in PDAC. The proposed research is significant because it is expected to advance the fields of PDAC treatment and oxidative stress biology by establishing GSTP1 as a new therapeutic target for PDAC treatment and revealing novel cellular responses to inhibition of this important antioxidant enzyme. The results from this project are expected to expand understanding of how antioxidant enzymes can be targeted for chemosensitization of tumor cells. Such knowledge has the potential to change the field of chemotherapy and result in more effective cancer treatments. Given that treatment options for PDAC are currently lacking, there is an urgent need to identify safe and more effective treatment approaches for this highly deadly disease. Finally, this project will have a tremendous impact on the undergraduate and graduate students involved, and the biomedical research capacity of North Dakota State University.