Proteolytic Cleavage of Apolipoprotein E in the Down Syndrome Brain

Down syndrome (DS) is one of the most common genetic causes of intellectual disability and is characterized by a number of behavioral as well as cognitive symptoms. Many of the neuropathological features of early-onset Alzheimer’s disease (AD) including senile plaques and neurofibrillary tangles (NFTs) are also present in people with DS as a result of triplication of the amyloid precursor gene on chromosome 21. Evidence suggests that harboring one or both apolipoprotein E4 (APOE4) alleles may increase the risk for AD due to the proteolytic cleavage of apoE4 and a subsequent loss of function. To investigate a role for the apoE proteolysis in vivo, we compared three autopsy groups; 7 DS with AD neuropathology cases over 40 years, 5 young DS cases without AD pathology under 40 years (YDS) and 5 age-matched control cases over 40 years by immunohistochemistry utilizing an antibody that detects the amino-terminal fragment of apoE. Application of this antibody, termed the amino-terminal apoE fragment antibody (nApoECF) revealed labeling of pyramidal neurons in the frontal cortex of YDS cases, whereas in the DS-AD group, labeling with nApoECF was prominent within NFTs. NFT labeling with nApoECF was significantly greater in the hippocampus versus the frontal cortex in the same DS-AD cases, suggesting a regional distribution of truncated apoE. Colocalization immunofluorescence experiments indicated that 52.5% and 53.2% of AT8- and PHF-1-positive NFTs, respectively, also contained nApoECF. Collectively, these data support a role for the proteolytic cleavage of apoE in DS and suggest that apoE fragmentation is closely associated with NFTs

Generating Stable Bioluminescent Cell Lines by Retroviral Transduction

This project focuses on improving tumor in vivo imaging techniques. It is difficult to accurately in vivo image orthotopic tumors using CT scans alone. Muriglo provides the capability to visualize in vivo bioluminescent signals emanating from intact tissues. We can coregister CT scans captured with the SARRP with luciferase signals captured with the Muriglo and locate tumor isocenters for image-guided radiation. We may then direct radiation to neoplastic tissue while also sparing the maximum amount of healthy tissue. To do this we must genetically engineer cells to express luciferase. We propose a protocol to use a retrovirus to create stable cell lines that express firefly luciferase for bioluminescence. We will also determine the optimal dosage of the G418 antibiotic for selection

Promoting Inclusion of Adults with Disabilities in Local Fitness Programs: A Needs Assessment

Fit-Pals* is a university-based, service-learning program with a mission to prepare adults with disabilities to engage in lifelong physical activity. We conducted a Needs Assessment to evaluate recent programmatic partnerships with community-based fitness organizations. We aimed to (1) evaluate organizational perceptions of Fit-Pals’ partnership efforts, and (2) identify perceived organizational needs to improve inclusion practices. Representatives from each of our seven partner organizations participated in an online-survey, follow-up interviews, and a stakeholder meeting. A thematic analysis of survey and interview responses highlighted areas for programmatic growth related to training in disability awareness and fitness accommodations, and improved communication across all partnership levels. Our stakeholder meeting further identified gaps between advocacy for disability inclusion, and awareness of actionable steps to effectively enact this within organizations. Drawing from the literature we discuss Fit-Pals’ efforts to increase the capacity of our community partners to support members with disabilities. *Pseudony

Efficacy of Service Dogs with Veterans with Post-Traumatic Stress Disorder (PTSD): Community Reintegration

Purpose: The purpose of this study was to determine if having a service dog is effective for improving community reintegration (social aspects, mental/physical health, safety, companionship, employment, quality of life) of veterans with post-traumatic stress disorder (PTSD). Methods: In this longitudinal pre-and post-treatment mixed-methods study, participants received a survey prior to receiving their service dog to obtain baseline data of various aspects of occupational engagement and reintegration into the community. After receiving their service dogs post surveys were completed at 3, 6, 9, and 12 months. Quantitative results were analyzed using descriptive statistics and compared to the baseline to determine changes and qualitative data was used to determine common themes through content analysis. Results: The study recruited 23 participants however, many participants did not provide multiple data sets impeding evaluation of the impact of a service dog over time for community reintegration. Data analysis was adjusted and organized according to trends of relevant responses over time. Qualitative data was analyzed and compared to trends as well. Conclusion: The current researchers found a positive correlation between veterans and service dogs while reintegrating into the community. Service dogs provided the veterans with an increase in confidence, social participation, self-care, emotional regulation, comfortability while entering back into the community, and decreased levels of stress

Blockade of fibroblast activation protein in combination with radiation treatment in murine models of pancreatic adenocarcinoma

Pancreatic ductal adenocarcinoma (PDAC) is characterized by a fibrotic stroma with a poor lymphocyte infiltrate, in part driven by cancer-associated fibroblasts (CAFs). CAFs, which express fibroblast activation protein (FAP), contribute to immune escape via exclusion of anti-tumor CD8+ T cells from cancer cells, upregulation of immune checkpoint ligand expression, immunosuppressive cytokine production, and polarization of tumor infiltrating inflammatory cells. FAP is a post-proline peptidase selectively expressed during tissue remodeling and repair, such as with wound healing, and in the tumor microenvironment by cancer-associated fibroblasts. We targeted FAP function using a novel small molecule inhibitor, UAMC-1110, and mice with germline knockout of FAP and concomitant knock-in of E. coli beta-galactosidase. We depleted CAFs by adoptive transfer of anti-βgal T cells into the FAP knockout animals. Established syngeneic pancreatic tumors in immune competent mice were targeted with these 3 strategies, followed by focal radiotherapy to the tumor. FAP loss was associated with improved antigen-specific tumor T cell infiltrate and enhanced collagen deposition. However, FAP targeting alone or with tumor-directed radiation did not improve survival even when combined with anti-PD1 therapy. Targeting of CAFs alone or in combination with radiation did not improve survival. We conclude that targeting FAP and CAFs in combination with radiation is capable of enhancing anti-tumor T cell infiltrate and function, but does not result in sufficient tumor clearance to extend survival.</div