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Aggregates of medin amyloid (a fragment of the protein MFG-E8, also known as lactadherin) are found in the vasculature of almost all humans over 50 years of age(1,)(2), making it the most common amyloid currently known. We recently reported that medin also aggregates in blood vessels of ageing wild-type mice, causing cerebrovascular dysfunction(3). Here we demonstrate in amyloid-beta precursor protein (APP) transgenic mice and in patients with Alzheimers disease that medin co-localizes with vascular amyloid-beta deposits, and that in mice, medin deficiency reduces vascular amyloid-beta deposition by half. Moreover, in both the mouse and human brain, MFG-E8 is highly enriched in the vasculature and both MFG-E8 and medin levels increase with the severity of vascular amyloid-beta burden. Additionally, analysing data from 566 individuals in the ROSMAP cohort, we find that patients with Alzheimers disease have higher MFGE8 expression levels, which are attributable to vascular cells and are associated with increased measures of cognitive decline, independent of plaque and tau pathology. Mechanistically, we demonstrate that medin interacts directly with amyloid-beta to promote its aggregation, as medin forms heterologous fibrils with amyloid-beta, affects amyloid-beta fibril structure, and cross-seeds amyloid-beta aggregation both in vitro and in vivo. Thus, medin could be a therapeutic target for prevention of vascular damage and cognitive decline resulting from amyloid-beta deposition in the blood vessels of the brain., Funding Agencies|Deutsche Forschungsgemeinschaft (DFG) [NE 1951/2-1, NE 1951/2-2, JU655/4-1]; EU/EFPIA/Innovative Medicines Initiative 2 Joint Undertaking (IMPRiND grant) [116060]; Hans und Ilse Breuer-Stiftung; China Scholarship Council; FEBS short term travel fellowship; British Heart Foundation Intermediate Basic Science fellowship [FS/12/61/29877]; Swedish Research Council; Flanders Institute for Biotechnology (VIB) [C0401]; Fund for Scientific Research Flanders (FWO) [G0C3522N, 12P0919N, 12P0922N]; Stichting Alzheimer Onderzoek/Fondation Recherche Alzheimer [SAO-FRA 2020/0013]; Alzheimers Research UK senior fellowship; Alexander von Humboldt Foundation research fellowship; NIA [P30AG10161, R01AG15819, R01AG17917, U01AG46152, U01AG61356, P50AG025688]; Illinois Department of Public Health (ROSMAP); Translational Genomics Research Institute

Aggregates of medin amyloid (a fragment of the protein MFG-E8, also known as lactadherin) are found in the vasculature of almost all humans over 50 years of age(1,)(2), making it the most common amyloid currently known. We recently reported that medin also aggregates in blood vessels of ageing wild-type mice, causing cerebrovascular dysfunction(3). Here we demonstrate in amyloid-beta precursor protein (APP) transgenic mice and in patients with Alzheimers disease that medin co-localizes with vascular amyloid-beta deposits, and that in mice, medin deficiency reduces vascular amyloid-beta deposition by half. Moreover, in both the mouse and human brain, MFG-E8 is highly enriched in the vasculature and both MFG-E8 and medin levels increase with the severity of vascular amyloid-beta burden. Additionally, analysing data from 566 individuals in the ROSMAP cohort, we find that patients with Alzheimers disease have higher MFGE8 expression levels, which are attributable to vascular cells and are associated with increased measures of cognitive decline, independent of plaque and tau pathology. Mechanistically, we demonstrate that medin interacts directly with amyloid-beta to promote its aggregation, as medin forms heterologous fibrils with amyloid-beta, affects amyloid-beta fibril structure, and cross-seeds amyloid-beta aggregation both in vitro and in vivo. Thus, medin could be a therapeutic target for prevention of vascular damage and cognitive decline resulting from amyloid-beta deposition in the blood vessels of the brain., Funding Agencies|Deutsche Forschungsgemeinschaft (DFG) [NE 1951/2-1, NE 1951/2-2, JU655/4-1]; EU/EFPIA/Innovative Medicines Initiative 2 Joint Undertaking (IMPRiND grant) [116060]; Hans und Ilse Breuer-Stiftung; China Scholarship Council; FEBS short term travel fellowship; British Heart Foundation Intermediate Basic Science fellowship [FS/12/61/29877]; Swedish Research Council; Flanders Institute for Biotechnology (VIB) [C0401]; Fund for Scientific Research Flanders (FWO) [G0C3522N, 12P0919N, 12P0922N]; Stichting Alzheimer Onderzoek/Fondation Recherche Alzheimer [SAO-FRA 2020/0013]; Alzheimers Research UK senior fellowship; Alexander von Humboldt Foundation research fellowship; NIA [P30AG10161, R01AG15819, R01AG17917, U01AG46152, U01AG61356, P50AG025688]; Illinois Department of Public Health (ROSMAP); Translational Genomics Research Institute

Aggregates of medin amyloid (a fragment of the protein MFG-E8, also known as lactadherin) are found in the vasculature of almost all humans over 50 years of age(1,)(2), making it the most common amyloid currently known. We recently reported that medin also aggregates in blood vessels of ageing wild-type mice, causing cerebrovascular dysfunction(3). Here we demonstrate in amyloid-beta precursor protein (APP) transgenic mice and in patients with Alzheimers disease that medin co-localizes with vascular amyloid-beta deposits, and that in mice, medin deficiency reduces vascular amyloid-beta deposition by half. Moreover, in both the mouse and human brain, MFG-E8 is highly enriched in the vasculature and both MFG-E8 and medin levels increase with the severity of vascular amyloid-beta burden. Additionally, analysing data from 566 individuals in the ROSMAP cohort, we find that patients with Alzheimers disease have higher MFGE8 expression levels, which are attributable to vascular cells and are associated with increased measures of cognitive decline, independent of plaque and tau pathology. Mechanistically, we demonstrate that medin interacts directly with amyloid-beta to promote its aggregation, as medin forms heterologous fibrils with amyloid-beta, affects amyloid-beta fibril structure, and cross-seeds amyloid-beta aggregation both in vitro and in vivo. Thus, medin could be a therapeutic target for prevention of vascular damage and cognitive decline resulting from amyloid-beta deposition in the blood vessels of the brain., Funding Agencies|Deutsche Forschungsgemeinschaft (DFG) [NE 1951/2-1, NE 1951/2-2, JU655/4-1]; EU/EFPIA/Innovative Medicines Initiative 2 Joint Undertaking (IMPRiND grant) [116060]; Hans und Ilse Breuer-Stiftung; China Scholarship Council; FEBS short term travel fellowship; British Heart Foundation Intermediate Basic Science fellowship [FS/12/61/29877]; Swedish Research Council; Flanders Institute for Biotechnology (VIB) [C0401]; Fund for Scientific Research Flanders (FWO) [G0C3522N, 12P0919N, 12P0922N]; Stichting Alzheimer Onderzoek/Fondation Recherche Alzheimer [SAO-FRA 2020/0013]; Alzheimers Research UK senior fellowship; Alexander von Humboldt Foundation research fellowship; NIA [P30AG10161, R01AG15819, R01AG17917, U01AG46152, U01AG61356, P50AG025688]; Illinois Department of Public Health (ROSMAP); Translational Genomics Research Institute

Aggregates of medin amyloid (a fragment of the protein MFG-E8, also known as lactadherin) are found in the vasculature of almost all humans over 50 years of age(1,)(2), making it the most common amyloid currently known. We recently reported that medin also aggregates in blood vessels of ageing wild-type mice, causing cerebrovascular dysfunction(3). Here we demonstrate in amyloid-beta precursor protein (APP) transgenic mice and in patients with Alzheimers disease that medin co-localizes with vascular amyloid-beta deposits, and that in mice, medin deficiency reduces vascular amyloid-beta deposition by half. Moreover, in both the mouse and human brain, MFG-E8 is highly enriched in the vasculature and both MFG-E8 and medin levels increase with the severity of vascular amyloid-beta burden. Additionally, analysing data from 566 individuals in the ROSMAP cohort, we find that patients with Alzheimers disease have higher MFGE8 expression levels, which are attributable to vascular cells and are associated with increased measures of cognitive decline, independent of plaque and tau pathology. Mechanistically, we demonstrate that medin interacts directly with amyloid-beta to promote its aggregation, as medin forms heterologous fibrils with amyloid-beta, affects amyloid-beta fibril structure, and cross-seeds amyloid-beta aggregation both in vitro and in vivo. Thus, medin could be a therapeutic target for prevention of vascular damage and cognitive decline resulting from amyloid-beta deposition in the blood vessels of the brain., Funding Agencies|Deutsche Forschungsgemeinschaft (DFG) [NE 1951/2-1, NE 1951/2-2, JU655/4-1]; EU/EFPIA/Innovative Medicines Initiative 2 Joint Undertaking (IMPRiND grant) [116060]; Hans und Ilse Breuer-Stiftung; China Scholarship Council; FEBS short term travel fellowship; British Heart Foundation Intermediate Basic Science fellowship [FS/12/61/29877]; Swedish Research Council; Flanders Institute for Biotechnology (VIB) [C0401]; Fund for Scientific Research Flanders (FWO) [G0C3522N, 12P0919N, 12P0922N]; Stichting Alzheimer Onderzoek/Fondation Recherche Alzheimer [SAO-FRA 2020/0013]; Alzheimers Research UK senior fellowship; Alexander von Humboldt Foundation research fellowship; NIA [P30AG10161, R01AG15819, R01AG17917, U01AG46152, U01AG61356, P50AG025688]; Illinois Department of Public Health (ROSMAP); Translational Genomics Research Institute

Aggregates of medin amyloid (a fragment of the protein MFG-E8, also known as lactadherin) are found in the vasculature of almost all humans over 50 years of age(1,)(2), making it the most common amyloid currently known. We recently reported that medin also aggregates in blood vessels of ageing wild-type mice, causing cerebrovascular dysfunction(3). Here we demonstrate in amyloid-beta precursor protein (APP) transgenic mice and in patients with Alzheimers disease that medin co-localizes with vascular amyloid-beta deposits, and that in mice, medin deficiency reduces vascular amyloid-beta deposition by half. Moreover, in both the mouse and human brain, MFG-E8 is highly enriched in the vasculature and both MFG-E8 and medin levels increase with the severity of vascular amyloid-beta burden. Additionally, analysing data from 566 individuals in the ROSMAP cohort, we find that patients with Alzheimers disease have higher MFGE8 expression levels, which are attributable to vascular cells and are associated with increased measures of cognitive decline, independent of plaque and tau pathology. Mechanistically, we demonstrate that medin interacts directly with amyloid-beta to promote its aggregation, as medin forms heterologous fibrils with amyloid-beta, affects amyloid-beta fibril structure, and cross-seeds amyloid-beta aggregation both in vitro and in vivo. Thus, medin could be a therapeutic target for prevention of vascular damage and cognitive decline resulting from amyloid-beta deposition in the blood vessels of the brain., Funding Agencies|Deutsche Forschungsgemeinschaft (DFG) [NE 1951/2-1, NE 1951/2-2, JU655/4-1]; EU/EFPIA/Innovative Medicines Initiative 2 Joint Undertaking (IMPRiND grant) [116060]; Hans und Ilse Breuer-Stiftung; China Scholarship Council; FEBS short term travel fellowship; British Heart Foundation Intermediate Basic Science fellowship [FS/12/61/29877]; Swedish Research Council; Flanders Institute for Biotechnology (VIB) [C0401]; Fund for Scientific Research Flanders (FWO) [G0C3522N, 12P0919N, 12P0922N]; Stichting Alzheimer Onderzoek/Fondation Recherche Alzheimer [SAO-FRA 2020/0013]; Alzheimers Research UK senior fellowship; Alexander von Humboldt Foundation research fellowship; NIA [P30AG10161, R01AG15819, R01AG17917, U01AG46152, U01AG61356, P50AG025688]; Illinois Department of Public Health (ROSMAP); Translational Genomics Research Institute

© 2020 American Association for Cancer Research. Pancreatic ductal adenocarcinoma (PDAC) has a poor 5-year survival rate and lacks effective therapeutics. Therefore, it is of paramount importance to identify new targets. Using multiplex data from patient tissue, three-dimensional coculturing in vitro assays, and orthotopic murine models, we identified Netrin G1 (NetG1) as a promoter of PDAC tumorigenesis. We found that NetG1+ cancer-associated fibroblasts (CAF) support PDAC survival, through a NetG1-mediated effect on glutamate/glutamine metabolism. Also, NetG1+ CAFs are intrinsically immunosup-pressive and inhibit natural killer cell–mediated killing of tumor cells. These protumor functions are controlled by a signaling circuit downstream of NetG1, which is comprised of AKT/4E-BP1, p38/FRA1, vesicular glutamate transporter 1, and glutamine synthetase. Finally, blocking NetG1 with a neutral-izing antibody stunts in vivo tumorigenesis, suggesting NetG1 as potential target in PDAC. SIGNIFICANCE: This study demonstrates the feasibility of targeting a fibroblastic protein, NetG1, which can limit PDAC tumorigenesis in vivo by reverting the protumorigenic properties of CAFs. Moreover, inhibition of metabolic proteins in CAFs altered their immunosuppressive capacity, linking metabolism with immunomodulatory function.

© 2020 American Association for Cancer Research. Pancreatic ductal adenocarcinoma (PDAC) has a poor 5-year survival rate and lacks effective therapeutics. Therefore, it is of paramount importance to identify new targets. Using multiplex data from patient tissue, three-dimensional coculturing in vitro assays, and orthotopic murine models, we identified Netrin G1 (NetG1) as a promoter of PDAC tumorigenesis. We found that NetG1+ cancer-associated fibroblasts (CAF) support PDAC survival, through a NetG1-mediated effect on glutamate/glutamine metabolism. Also, NetG1+ CAFs are intrinsically immunosup-pressive and inhibit natural killer cell–mediated killing of tumor cells. These protumor functions are controlled by a signaling circuit downstream of NetG1, which is comprised of AKT/4E-BP1, p38/FRA1, vesicular glutamate transporter 1, and glutamine synthetase. Finally, blocking NetG1 with a neutral-izing antibody stunts in vivo tumorigenesis, suggesting NetG1 as potential target in PDAC. SIGNIFICANCE: This study demonstrates the feasibility of targeting a fibroblastic protein, NetG1, which can limit PDAC tumorigenesis in vivo by reverting the protumorigenic properties of CAFs. Moreover, inhibition of metabolic proteins in CAFs altered their immunosuppressive capacity, linking metabolism with immunomodulatory function.

Oxidative stress has been implicated as a cause of the protein destabilization and cytoskeletal disruption that have been linked to the onset and progression of neurodegenerative disease. Findings that mixed disulfides between proteins and glutathione form in response to oxidative stress and that this modification (S-glutathionylation) may destabilize some proteins has prompted our hypothesis that unregulated glutathionylation of proteins could be a contributor to protein destabilization and cytoskeletal disruption in the brain. In this study, we examined using biotinyl-glutathione and avidin-affinity chromatography the presumed non-enzymatic glutathionylation of rat brain proteins, with a particular interest in the cytoskeletal proteins tubulin and actin. In addition, we explored in preliminary experiments the possibility that a rapid glutathione-induced destabilization in vitro of total soluble proteins from rat brain may involve glutathionylation and, perhaps, an interaction between glutathione and either tubulin or actin. Our results demonstrate that tubulin and actin are subject to substantial glutathionylation in vitro by an apparent nonenzymatic pathway. While the basis of the glutathione-induced destabilization of brain proteins remains unknown, our results suggest that this response may be associated with the presence of tubulin but not with actin. Overall, these findings support a complex interaction between glutathione and brain proteins.

Transcranial direct current stimulation (tDCS) has been shown to modulate neuroplasticity. Beneficial effects are observed in patients with psychiatric disorders and enhancement of brain performance in healthy individuals has been observed following tDCS. However, few studies have attempted to elucidate the underlying molecular mechanisms of tDCS in the brain. This study was conducted to assess the impact of tDCS on gene expression within the rat cerebral cortex. Anodal tDCS was applied at 3 different intensities followed by RNA-sequencing and analysis. In each current intensity, approximately 1,000 genes demonstrated statistically significant differences compared to the sham group. A variety of functional pathways, biological processes, and molecular categories were found to be modified by tDCS. The impact of tDCS on gene expression was dependent on current intensity. Results show that inflammatory pathways, antidepressant-related pathways (GTP signaling, calcium ion binding, and transmembrane/signal peptide pathways), and receptor signaling pathways (serotonergic, adrenergic, GABAergic, dopaminergic, and glutamate) were most affected. Of the gene expression profiles induced by tDCS, some changes were observed across multiple current intensities while other changes were unique to a single stimulation intensity. This study demonstrates that tDCS can modify the expression profile of various genes in the cerebral cortex and that these tDCS-induced alterations are dependent on the current intensity applied.

Transcranial direct current stimulation (tDCS) has been shown to modulate neuroplasticity. Beneficial effects are observed in patients with psychiatric disorders and enhancement of brain performance in healthy individuals has been observed following tDCS. However, few studies have attempted to elucidate the underlying molecular mechanisms of tDCS in the brain. This study was conducted to assess the impact of tDCS on gene expression within the rat cerebral cortex. Anodal tDCS was applied at 3 different intensities followed by RNA-sequencing and analysis. In each current intensity, approximately 1,000 genes demonstrated statistically significant differences compared to the sham group. A variety of functional pathways, biological processes, and molecular categories were found to be modified by tDCS. The impact of tDCS on gene expression was dependent on current intensity. Results show that inflammatory pathways, antidepressant-related pathways (GTP signaling, calcium ion binding, and transmembrane/signal peptide pathways), and receptor signaling pathways (serotonergic, adrenergic, GABAergic, dopaminergic, and glutamate) were most affected. Of the gene expression profiles induced by tDCS, some changes were observed across multiple current intensities while other changes were unique to a single stimulation intensity. This study demonstrates that tDCS can modify the expression profile of various genes in the cerebral cortex and that these tDCS-induced alterations are dependent on the current intensity applied.

Transcranial direct current stimulation (tDCS) has been shown to modulate neuroplasticity. Beneficial effects are observed in patients with psychiatric disorders and enhancement of brain performance in healthy individuals has been observed following tDCS. However, few studies have attempted to elucidate the underlying molecular mechanisms of tDCS in the brain. This study was conducted to assess the impact of tDCS on gene expression within the rat cerebral cortex. Anodal tDCS was applied at 3 different intensities followed by RNA-sequencing and analysis. In each current intensity, approximately 1,000 genes demonstrated statistically significant differences compared to the sham group. A variety of functional pathways, biological processes, and molecular categories were found to be modified by tDCS. The impact of tDCS on gene expression was dependent on current intensity. Results show that inflammatory pathways, antidepressant-related pathways (GTP signaling, calcium ion binding, and transmembrane/signal peptide pathways), and receptor signaling pathways (serotonergic, adrenergic, GABAergic, dopaminergic, and glutamate) were most affected. Of the gene expression profiles induced by tDCS, some changes were observed across multiple current intensities while other changes were unique to a single stimulation intensity. This study demonstrates that tDCS can modify the expression profile of various genes in the cerebral cortex and that these tDCS-induced alterations are dependent on the current intensity applied.

This project aimed to investigate the molecular mechanism(s) of organ injury following exposure to sublethal doses of the organophosphorus agent, sarin. The lethal toxic effect of sarin exposure is caused by irreversible inhibition of acetylcholinesterase (AChE) and subsequently excessive stimulation of postsynaptic cholinergic receptors. However, much less is known about the effects of low-dose sarin exposure on functions of critical brain regions. Neurotoxicity caused by sarin exposure can be mediated by pathways independent of the cholinergic system. Since the hippocampus plays an important role in cognitive functions (including learning and memory), and is one of the most sensitive brain regions to sarin-induced injury, the focus of this study is to investigate differential gene expression and pathway perturbation in this brain region, using microarray technology combined with advanced bioinformatics. Sarin treatment resulted in differential expression of a large number of genes. Most of these genes showed increased expression, especially at the 6-hour and 25-day time points. However, significant gene repression was observed at the 10-day time point. Pathway analysis revealed that one of the direct effects of sarin cellular toxicity could be related to DNA damage and inhibition of the glutathione antioxidant system, which triggered the up-regulation of the apoptotic pathways at early time points., Prepared in collaboration with the Molecular Bioeffects Branch, Human Effectiveness Directorate, Air Force Research Laboratory, Wright-Patterson AFB OH. This project was funded by Defense Threats Reduction Agency (DTRA). Employees of Henry M. Jackson Foundation (HJF) were working under the Cooperative Agreement FA8650-05-2-6518 between RHDJ and HJF. Animal exposure was conducted at Wright State University (WSU) under the HJF Cooperative Agreement FA8650-05-2-6518, Subaward # 667357.

This seedling project aimed to genetically map intelligence in the military working dog (MWD) population. A total of 199 canine subjects were recruited from United States working dog contractors. Of the recruited subjects, 153 were tested using the Canine Intelligence Testing Protocol (CITP), developed by Dr. Karen Overall (UPENN) to specifically analyze canine intelligence. CITP allows quantitative assessment of intelligence in individual dogs using a scoring system based on the latency to response, success-in-effort time, attentiveness, interest in novelty exploration, response to signaling and showing, observational learning, problem solving/boldness, and handedness. Blood samples were collected from the canines, and genomic DNA prepared. A total of 117 dogs, belonging to three breeds (German Shepherds, Belgian Malinois, Labrador Retrievers) were down-selected and successfully genotyped for whole genome (WG) single nucleotide polymorphism (SNP) markers by means of the Affymetrix Canine SNP Array v2. A proof-of-concept advanced data mining algorithm for unsupervised analysis of the genome-wide association study (GWAS) dataset was successfully developed. Using this algorithm, canine subjects were successfully clustered into the correct breeds with an accuracy ranging from 89% - 100%, solely based on the WG SNP profiles. The details of the algorithm are described in the Technical Report AFRL-RH-WP-TR-2011-0081, entitled Development of Advanced Classification Algorithm for Genome-Wide Single Nucleotide Polymorphism (SNP) Data Analysis. While not initially part of the seedling proposal, this project did receive IACUC permission to test DoD MWDs for follow-on studies, a unique and significant accomplishment., Sponsored in part by DARPA. The original document contains color images.

This project collected data to genetically map superior intelligence in the military working dog. A behavioral testing regimen was developed by canine cognitive expert Dr Karen Overall (UPENN) which enabled quantitative intelligence testing of individual dogs and blood samples were taken, and genome-wide SNP typing completed by means of the Affymetrix Canine SNP (single nucleotide polymorphism) Array v2. In order to identify SNP markers for mapping of small-effect-sized genes that contribute to highly complex polygenic traits, it is necessary to develop a more robust computational method for the analysis of SNP profile data. To accomplish this, we are undertaking two parallel efforts, Biologically Guided Feature Selection and Computational Based Feature Synthesis and Classification. As a proof-of-concept, we conducted a classification analysis focused on a subset of tested canines consisting of German Shepherds, Labrador Retrievers, and Belgian Malinois. Using this new classification technique, samples from the three breeds clustered into the correct breed with an accuracy ranging from 89 - 100 %. Classification accuracy was not significantly affected by data process methods (including data cleanup methods) or SNP annotation quality, thus suggesting that this algorithm is highly robust. With further refinement and optimization, this technique could be used to classify complex phenotypes in an unsupervised manner and allow identification of associated SNP markers., Prepared in cooperation with Penn Med Translation Research Lab., Philadelphia, PA, Ohio Univ., School of Electrical Engineering and Computer Science

This paper will try to untangle how the U.S. Office of War Information's Bureau of Motion Pictures tried to enact change in the world, using Hollywood films, during World War II. It will also show how inconsistencies within the agency and lack of support from the President, Congress, and Hollywood often sabotaged the Bureau's project. I argue that a structural component and a thematic component helped cripple the OWI's Bureau of Motion Pictures. First, the extremely decentralized, bureaucratic and conflict-laden nature of the government information network, and the limited enforcement power of the OWI and in particular the Bureau of Motion Pictures, limited its success. Second, the BMP's passionately liberal and racially progressive interpretation of U.S. war aims helped contribute to its downfall. The BMP operated during a watershed moment in race relations, in which hierarchies of racial and ethnic groups were shifting dramatically.