Marc D. Basson - US grants

DESCRIPTION (Verbatim from Applicant's Abstract): Cytoskeletal-associated signal proteins and their downstream effectors are a common pathway for signal transduction from the extracellular matrix, soluble growth factors and physical forces, such as strain and shear stress. The effects of forces generated by increases in extracellular pressure are less well understood but may act via similar pathways. The classic pathway for initiating outside-in signaling is through the binding of cells to extracellular matrix proteins via various integrins. The mechanisms through which intracellular events alter integrin-mediated cell adhesion to matrix proteins, inside-out signaling, are less well understood. Preliminary studies in four colon cancer cell lines and primary cells from 12 human colon tumors suggest that increases in ambient pressure as low as 10mmHg stimulate colon cancer cell, cation dependant adhesiveness. This occurs synergistically with an adhesion promoting factor in serum. Based on preliminary data and what is known about physical force transduction and inside-out signaling in other cell types, the authors hypothesize that increased extracellular pressure initiates a cytoskeletally associated tyrosine kinase mediated signal transduction cascade involving focal adhesion kinase and members of the MAPK family which alters malignant colonocyte integrin binding. This proposal seeks to further define the mechanism by which pressure stimulates adhesion. The authors will identify agents that prevent pressure-stimulated tumor cell adhesion. In the study of these FAK and MAPK enzymes, the basis for a new pathway which regulates cell adhesiveness may be found.

DESCRIPTION (provided by applicant): Trauma and burns are the leading cause of death and morbidity in young individuals. Despite modest advances in the early care of these patients, complications and mortality associated with hemorrhage, shock, tissue injury, infection, wound healing and multi system organ failure remain excessive. Continued improvements in patient outcomes will require fundamental research to identify novel treatment strategies. The objective of the proposed program is to identify and train young surgeons for continuing, productive academic careers focusing of problems relating to trauma and burns. The program insures that individuals possess the necessary basic science and clinical research skills as well as the environment to successfully both identify and subsequently investigate key research questions that will affect the understanding and management of problems affecting trauma and burn victims. Clinical advances in trauma and burn care require not only careful definition of current problems and deliberate basic investigations of ongoing pathophysiologic questions, but also application of leading edge methodologies in order to provide meaningful answers. The proposed program exposes surgical trainees to state of the art basic science research techniques and strategies to allow future academic success. This proposal involves 3 trainees/year who are general surgery residents and will spend a minimum of two years doing basic research.

DESCRIPTION (provided by applicant): Cancer cell attachment to distant tissues is fundamental to metastasis. We propose that signals within tumor cells interact with the cytoskeleton to modify the adhesiveness of cell surface receptors required for adhesion and metastasis. Based on preliminary data using established cell lines, a murine model, and primary cells from human colon cancers, we hypothesize that inside-out regulation of integrin receptors governs cancer cell adhesion. In particular, we have shown that increased extracellular pressure, such as may occur during passage through the venous or lymphatic system, during surgical procedures, or postoperatively due to increased intra-abdominal pressure, stimulates FAK-Src interaction and activates PI3-K together with the PI3-K substrate Akt. Preliminary studies suggest that these signals influence cancer cell adhesion in a manner that requires an intact actin cytoskeleton. We will now use pressure activation to further define the mechanisms of inside-out regulation of cancer cell adhesion. We will delineate the signals involved, explore the role of the cytoskeleton, and determine whether adhesiveness is regulated by changes in integrin affinity or avidity by measuring both integrin clustering and the bond strength of individual integrin-matrix bonds in cancer cells subjected to pressure. We will test our hypothesis in human primary cancer cells and in a mouse model, to establish its relevance to intact organisms. This work will elucidate a scientifically important paradigm for inside-out signaling that may explain why some circulating cancer cells may adhere while others do not. The signals we define will represent important targets for perioperative intervention to reduce iatrogenic tumor spread during surgery and perhaps new targets for cancer therapy to inhibit metastasis.

DESCRIPTION (provided by applicant): Mucosal healing requires epithelial motility, proliferation, and differentiation. Focal adhesion kinase (FAK) influences all three. FAK is an autophosphorylating tyrosine kinase that mediates downstream signals by receptors for matrix proteins and many growth factors and may promote epithelial cell motility, a critical first step in mucosal healing. Most studies have focused on FAK activation within minutes after ligand binding to FAK-associated membrane receptors. However, our preliminary studies suggest that FAK may be regulated at the protein level as well as in its phosphorylation during gut epithelial cell motility in vitro and during mucosal healing in vivo. This grant will focus on the exploration of the regulation of FAK protein levels, about which little is known. We hypothesize that integrin- and FAK- related signal events regulate FAK protein pools at the mRNA level, by modulating FAK gene transcription or FAK mRNA degradation, and that TGFbeta stimulates FAK by acting on this pathway. We now propose to demonstrate that FAK protein changes are biologically significant, identify the key control points for regulation of intracellular FAK protein and mRNA pools during intestinal epithelial motility, and determine how TGFbeta regulates FAK protein and mRNA in migrating intestinal epithelial cells and what signals mediate these effects. We will validate our in vitro observations in a murine Smad 3 knockout mouse model and in tissue from human mucosal wounds, including peptic ulcers and inflammatory bowel disease. We expect to demonstrate the importance of protein levels of this critical molecule in gut epithelial wound healing and to define the mechanisms that regulate its synthesis. These results will characterize a previously unknown pathway by which intestinal epithelial cell motility is regulated and its specific modulation by TGFbeta. We can then, in the future, study how this pathway goes awry in instances of defective mucosal healing and target pharmacologic interventions to improve mucosal healing. Furthermore, although different cells may exhibit different patterns of FAK regulation, this work will also provide a paradigm by which FAK protein regulation can be studied in other cells and settings, where FAK may be involved in processes as diverse as malignant transformation and atherogenesis.

DESCRIPTION (provided by applicant): Mucosal atrophy often leads to barrier failure and sepsis in starving or fasting patients in spite of total parenteral nutrition (TPN). In addition, mny patients who undergo massive bowel resection fail to adapt and must be maintained on permanent TPN with high mortality. Current therapies aimed at preventing mucosal atrophy and enhancing adaptation are limited in efficacy, perhaps because they mostly induce proliferation. We propose that we must learn to promote differentiation as well as proliferation to achieve an intact functional mucosa, reverse mucosal atrophy, and engender maximal intestinal adaptation. Some Schlafen-family proteins mediate cell growth, differentiation, or development in some cells in a Schlafen- and cell-specific fashion. We have demonstrated that the rodent protein Schlafen-3 (Slfn3) is induced during and required for intestinal epithelial differentiation in response to various stimuli. Our data indicate that Slfn3 varies with mucosal atrophy and promotes differentiation in vivo and that Schlafen-12 (SLFN12) is its human ortholog. We hypothesize that Schlafen 3/12 induction represents a fundamental and essential common pathway for enterocytic differentiation that can be specifically targeted to promote differentiation and maintain the mucosa. We further propose that Schlafens 3/12 are chaperoned to the nucleus where they enhance differentiation by affecting transcription and by a Cdx2 homeobox protein- dependent positive feedback mechanism. We will study intestinal mucosa in patients fasting on TPN (atrophy) or after Roux-en-Y gastric bypass (hypertrophy), rat and human intestinal epithelial cells in vitro, a novel rodent model of in vivo intestinal epithelial atrophy and hypertrophy, in vivo viral Slfn3/SLFN12 infection, and a tet-inducible intestine-specific Slfn3 knock-out mouse to achieve the following specific aims: 1) show that SLFN12 mediates human enterocytic differentiation and mirrors Slfn3 changes with differentiation in mucosal atrophy or hypertophy, 2) determine the mechanism by which Slfn3 and SLFN12 promote differentiation in rat and human intestinal epithelial cells, and 3) demonstrate that manipulating Slfn3/SLFN12 or a nuclear chaperone modulates intestinal atrophy/adaptation in vivo. This study will fill a critica knowledge gap because we will delineate a fundamental mechanism by which diverse stimuli regulate intestinal epithelial differentiation. This may facilitate interventions to maintain the mucosal barrier in starving or critically ill patients with mucosal atrophy and promote nutrition i pediatric or adult short bowel syndrome. The Slfn3 knock-out mouse will provide valuable insights not only into the convergent differentiation regulatory pathway we will study but also int normal gut development, mucosal healing and other intestinal disorders such as IBD. In addition, understanding how Schlafen proteins promote differentiation will have broader implications for differentiation and development in other epithelial tissues.

DESCRIPTION (provided by applicant): Mucosal atrophy often leads to barrier failure and sepsis in starving or fasting patients despite total parenteral nutrition (TPN). Many patients who undergo massive bowel resection fail to adapt and require permanent TPN with high mortality. Current interventions to prevent mucosal atrophy or enhance adaptation have limited efficacy, perhaps because they mostly induce proliferation. We hypothesize that hyperproliferation can be maladaptive as it is often accompanied by decreased differentiation. Thus, we propose that we must learn to promote differentiation as well as proliferation to achieve an intact functional mucosa, reverse mucosal atrophy, and engender maximal intestinal adaptation. Schlafen (Slfn)-family proteins mediate cell growth, differentiation, or development in hematopoietic cells in a Slfn and cell-specific fashion. We showed that the rodent protein Slfn3 is induced during and required for intestinal epithelial differentiation in response to various stimuli. We further found that Slfn3 varies with intestinal development and mucosal atrophy and promotes differentiation in vivo and that SLFN12 is its human ortholog. We hypothesize that Slfn3/12 induction represents a fundamental and essential common pathway for enterocytic differentiation that can be specifically targeted to promote differentiation and maintain the mucosa. We further propose that Slfns act in the cytosol through Slfn P-loop domain binding to specific proteins lowering proteasomal activity, increasing Cdx2 homeobox protein levels and enhancing transcription via a Cdx2-dependent positive feedback mechanism. We will study 1) intestinal mucosa in patients fasting on TPN (atrophy) or after Roux-en-Y gastric bypass (hypertrophy), 2) rat and human intestinal epithelial cells in vitro, and 3) the effects of in vivo manipulation of Slfn3/12 levelsin Slfn3 knock-out mice after 60% massive small bowel resection and a novel rodent model of intestinal epithelial atrophy and hypertrophy to: 1) Demonstrate that Slfn3 determines enterocytic differentiation in mucosal atrophy and adaptation in rodents, 2) Show that SLFN12 is the relevant human ortholog of Slfn3, and 3) Establish the mechanism(s) by which Slfn3/SLFN12 act. This study will fill a critical knowledge gap because we will delineate a fundamental mechanism by which diverse stimuli regulate intestinal epithelial differentiation. This will facilitate interventions to maintain the mucosal barrier in starving or critically ill paients with mucosal atrophy and promote nutrition in pediatric or adult short bowel syndrome. The Slfn3 knock-out mouse will provide valuable insights not only into the convergent differentiation regulatory pathway we will study but also into normal gut development, mucosal healing and other intestinal disorders such as IBD. In addition, understanding how Slfn proteins promote differentiation will have broader implications for differentiation and development in other epithelial tissues.

DACCOTA-Overall Summary Summary: The objective of this grant application is to create a center to support and expand our ability to conduct full-spectrum clinical and translational research on various cancers. Establishing the Dakota Cancer Collaborative on Translational Activity (DACCOTA) will also allow us for the first time to apply our basic scientific and epidemiologic work to the clinical arena while training clinician-investigator partners to translate these discoveries. We will train basic scientists to conduct translational work, population health experts to understand the community determinants of cancer outcomes, and clinicians to collaborate. The DACCOTA will promote and facilitate interactions between clinicians and scientists with unique but complementary areas of expertise. Our goal is to develop a highly productive, collaborative, and sustainable translational research center that will focus on the cancers that most commonly and disproportionately afflict the citizens of our region, especially American Indians. We have proposed independent cores, all led by capable investigators, that will assist in building a competitive clinical and translational cancer research center. The DACCOTA will provide an academic home for clinical and translational scientists and trainees. These investigators will focus on understanding the mechanisms leading to the initiation and progression of cancer, which will facilitate the development of novel and effective treatments and improve disease surveillance. This is necessary due to the increase in cancer prevalence and mortality in the Dakota region and throughout the USA. The DACCOTA will have further impact by validating new ways of developing and supporting individuals to facilitate team-based science addressing critical issues affecting the region. Our Professional Development Core will develop aspiring investigators and train practicing real world? clinicians to become more avid and effective collaborators. Applicants for pilot grants will be required to propose teams including an investigator and clinician. The Community Enagement and Outreach Core will help recruit volunteers, seek advice about overall directions, and work with community leaders to designate specific cancer-related issues of community concern for RFA?s that will be funneled through the Pilot Projects Program. In addition to the community- based natures of the University of North Dakota (UND), North Dakota State University (NDSU), and the University of South Dakota (USD), the collaborating hospitals in this proposal cover more than 90% of the region and include multiple innovative departments of family and community medicine as well as rural and population health. There are also three MPH programs between these institutions. Thus, we are superbly positioned to conduct clinical research encompassing the entire region. Similarly, the Biostatistics, Epidemiology, and Research Design Core and the Clinical Research Resources and Facilites Core will combine their strengths and develop new resources to enhance clinical research. Finally, our Tracking and Evaluation Core will use innovative methods to evaluate these novel processes as well as provide suggestions to improve efficiency and ensure both cost-effectiveness and improved patient safety. UND, NDSU, USD, and all clinical partners are fully committed to ensuring the growth and sustainability of the DACCOTA even after the CTR grant ends.

DACCOTA-Administrative Core Summary: The objective of the Adminstrative Core is to centralize the resources necessary to ensure the success of the Dakota Cancer Collaborative on Translational Activity (DACCOTA). This core will create integrated organizational governance to engage participating institutions in promoting the vision of the DACCOTA. Members of our center will function as a team rather than as individuals to achieve our objectives, with our guiding principle being the search for benefit for our patients. The Administrative Core will provide leadership and an administrative structure that supports day-to-day operations, coordinates, and oversees efforts to achieve the overall goals of the DACCOTA. This core will also foster collaborative engagement between partners and build an internal and external environment conducive to clinical and translational cancer research. To enhance the productivity of our clinical and translational investigators, this core will also leverage existing resources at the affiliated institutions as well as establish new cores and programs.

DACCOTA-Biostatistics, Epidemiology, and Research Design Core Summary: The Biostatistics, Epidemiology, and Research Design Core will provide support in the areas of methodology, biostatistics, epidemiology, research design, and biomedical informatics. We will create meaningful and enduring partnerships among the affiliated universities and research institutes to centralize and integrate methodological and bio-statistical research support for all participating clinicians and non-clinicians involved in cancer research. This core will promote solution?oriented studies and practice-based evidence to move evidence into action by bringing together a diverse group of professionals with expertise in the areas of methodology, biostatistics, epidemiology, and biomedical informatics related to cancer. This core will strengthen relationships across the policy, practice, and research sectors by building a high?performing and sustainable enterprise with a reputation for relevance, excellence, and influence. Our goal is to develop innovative biostatistics methods for translational science, including methods for solving diagnostic problems and evaluating causal mechanisms (such as environmental and genetic) that relate to cancer research. We also plan to develop new approaches to study recruitment that will improve the cost-effectiveness and efficiency of CTR.

DACCOTA-Clinical Research Resources and Facilities Core Summary: The overall goal of the Clinical Research Resources and Facilities Core is to provide synergy and expansion opportunities to the existing clinical services and technology cores available through partnering institutes, thereby fostering an ideal environment for the competitive execution of clinical and translational cancer research. This core will facilitate access to essential services necessary for establishing, executing, and communicating clinical and translational cancer research to allow for the seamless progression from ideas to results. The goal of this core is to promote CTR by expanding the biorepository of biological samples essential for advancing genomic/personalized medicine and translational research. Researchers will use these biobank samples to identify biomarkers for cancer progression and prognosis and/or determine medication tolerance. The DACCOTA will utilize an established biorepository with state-of-the-art technology to handle and inventory all samples, while also linking patient samples to the electronic medical records. The goal of this core is to share resources, develop new protocols for treating patients, and assist with various clinical trials as we move forward with our mission to increase clinical and translational cancer research.

DACCOTA-Pilot Projects Program Summary: The goal of the Pilot Projects Program is to provide seed funding for highly innovative projects addressing the needs of the local communities. The greatest hurdle in meeting this goal is the physical distance between our institutions, which minimizes the necessary familiarity of interests, skills, and goals critical for successful collaborations to proceed. Our institutions must also overcome cultural barriers that have resulted in historically limited insight into the research being conducted at other regional institutions. We will address this deficiency through an innovative online expertise and interest matching program to formalize interactions between our institutions and investigators. We will assemble teams of clinician and non-clinician investigators to promote the critical working relationship needed for a successful collaboration. Moreover, we propose three unique types of pilot grants stemming from the matching database. The first will be an innovative ?ready-to-go? pilot award for more mature clinician/non-clinician teams. The second will be pilot awards to clinicians/non-clinicians in response to specific solicitations generated through community input gathered by our Community Engagement and Outreach Core. The third pilot program is a feasibility award directed towards those clinician/non-clinician teams requiring additional input from the Pilot Projects Program and Biostatistics, Epidemiology and Research Design Core to formalize research design, statistical plans, and methodological needs in preparation for a larger pilot award. Using multiple funding mechanisms, we will foster collaborative translational research on topics ranging from molecules (genes/proteins) to populations and catalyze the translation of discoveries to treatments. Most importantly, all pilot awards will be derived from yearly participation in the interest/expertise matching software to improve the probability of success and provide a searchable database for tracking accomplishments and changes within our regional research effort.

Summary: The objective of this grant application is to create a center to support and expand our ability to conduct full-spectrum clinical and translational research on various cancers. Establishing the Dakota Cancer Collaborative on Translational Activity (DACCOTA) will also allow us for the first time to apply our basic scientific and epidemiologic work to the clinical arena while training clinician-investigator partners to translate these discoveries. We will train basic scientists to conduct translational work, population health experts to understand the community determinants of cancer outcomes, and clinicians to collaborate. The DACCOTA will promote and facilitate interactions between clinicians and scientists with unique but complementary areas of expertise. Our goal is to develop a highly productive, collaborative, and sustainable translational research center that will focus on the cancers that most commonly and disproportionately afflict the citizens of our region, especially American Indians. We have proposed independent cores, all led by capable investigators, that will assist in building a competitive clinical and translational cancer research center. The DACCOTA will provide an academic home for clinical and translational scientists and trainees. These investigators will focus on understanding the mechanisms leading to the initiation and progression of cancer, which will facilitate the development of novel and effective treatments and improve disease surveillance. This is necessary due to the increase in cancer prevalence and mortality in the Dakota region and throughout the USA. The DACCOTA will have further impact by validating new ways of developing and supporting individuals to facilitate team-based science addressing critical issues affecting the region. Our Professional Development Core will develop aspiring investigators and train practicing real world? clinicians to become more avid and effective collaborators. Applicants for pilot grants will be required to propose teams including an investigator and clinician. The Community Enagement and Outreach Core will help recruit volunteers, seek advice about overall directions, and work with community leaders to designate specific cancer-related issues of community concern for RFA?s that will be funneled through the Pilot Projects Program. In addition to the community-based natures of the University of North Dakota (UND), North Dakota State University (NDSU), and the University of South Dakota (USD), the collaborating hospitals in this proposal cover more than 90% of the region and include multiple innovative departments of family and community medicine as well as rural and population health. There are also three MPH programs between these institutions. Thus, we are superbly positioned to conduct clinical research encompassing the entire region. Similarly, the Biostatistics, Epidemiology, and Research Design Core and the Clinical Research Resources and Facilites Core will combine their strengths and develop new resources to enhance clinical research. Finally, our Tracking and Evaluation Core will use innovative methods to evaluate these novel processes as well as provide suggestions to improve efficiency and ensure both cost-effectiveness and improved patient safety. UND, NDSU, USD, and all clinical partners are fully committed to ensuring the growth and sustainability of the DACCOTA even after the CTR grant ends.

Summary: The objective of this grant application is to create a center to support and expand our ability to conduct full-spectrum clinical and translational research on various cancers. Establishing the Dakota Cancer Collaborative on Translational Activity (DACCOTA) will also allow us for the first time to apply our basic scientific and epidemiologic work to the clinical arena while training clinician-investigator partners to translate these discoveries. We will train basic scientists to conduct translational work, population health experts to understand the community determinants of cancer outcomes, and clinicians to collaborate. The DACCOTA will promote and facilitate interactions between clinicians and scientists with unique but complementary areas of expertise. Our goal is to develop a highly productive, collaborative, and sustainable translational research center that will focus on the cancers that most commonly and disproportionately afflict the citizens of our region, especially American Indians. We have proposed independent cores, all led by capable investigators, that will assist in building a competitive clinical and translational cancer research center. The DACCOTA will provide an academic home for clinical and translational scientists and trainees. These investigators will focus on understanding the mechanisms leading to the initiation and progression of cancer, which will facilitate the development of novel and effective treatments and improve disease surveillance. This is necessary due to the increase in cancer prevalence and mortality in the Dakota region and throughout the USA. The DACCOTA will have further impact by validating new ways of developing and supporting individuals to facilitate team-based science addressing critical issues affecting the region. Our Professional Development Core will develop aspiring investigators and train practicing real world? clinicians to become more avid and effective collaborators. Applicants for pilot grants will be required to propose teams including an investigator and clinician. The Community Enagement and Outreach Core will help recruit volunteers, seek advice about overall directions, and work with community leaders to designate specific cancer-related issues of community concern for RFA?s that will be funneled through the Pilot Projects Program. In addition to the community-based natures of the University of North Dakota (UND), North Dakota State University (NDSU), and the University of South Dakota (USD), the collaborating hospitals in this proposal cover more than 90% of the region and include multiple innovative departments of family and community medicine as well as rural and population health. There are also three MPH programs between these institutions. Thus, we are superbly positioned to conduct clinical research encompassing the entire region. Similarly, the Biostatistics, Epidemiology, and Research Design Core and the Clinical Research Resources and Facilites Core will combine their strengths and develop new resources to enhance clinical research. Finally, our Tracking and Evaluation Core will use innovative methods to evaluate these novel processes as well as provide suggestions to improve efficiency and ensure both cost-effectiveness and improved patient safety. UND, NDSU, USD, and all clinical partners are fully committed to ensuring the growth and sustainability of the DACCOTA even after the CTR grant ends.

DACCOTA-Tracking and Evaluation Core Summary: Efforts related to tracking, assessment, and evaluation of the various activities and programs of the DACCOTA will be conducted through the Tracking and Evaluation Core. This center will be an independent entity but will coordinate the Quality and Process Improvement Program by working closely with the Administrative Core. This independent evaluation represents a unique way in which the DACCOTA will leverage the expertise and resources of participating universities and institutions. The objective of this core is to examine and document the impact of the DACCOTA on the following: 1) promoting translational research across all phases of the research process, 2) creating a sustainable, efficient, cost-effective, and highly responsive environment for translational research, and 3) engaging diverse investigator communities to leverage these resources and emerge as a leader in cutting-edge translational research. This core?s analysis will inform the PI and administrative committees (including all core directors, the Internal Advisory Committee (IAC), and the External Advisory Committee (EAC)) on priorities for process improvement and initiatives to be implemented. Clear metrics of the success of the DACCOTA will be used to gauge performance across our partner institutions, programs, individuals, and communities. Success will be measured based on 1) promotion of interdisciplinary team science focused on clinical cancer research; 2) implementation of processes that enhance efficiency, outreach, quality, and safety in the full spectrum of translational research; 3) effective and efficient program management and implementation of changes as needed; and 4) development of effective clinical researchers who will thrive throughout their training, research initiatives, and career paths. Progress toward achieving these goals will be carefully monitored through a customized evaluation plan designed and administered by this core.