bluuebunny/crossref_metadata_2025_split huggingface dataset:
227-OR: Disruption of Adipocyte YAP Improves Glucose Homeostasis in Mice and Decreases Adipose Tissue Fibrosis
Authors: HAN DANIEL JONG JIN , ASLAM RUKHSANA , OJHA TANVI , YUEN DARREN , LUK CYNTHIA T. 
Published: May 2022

Adipose tissue is a dynamic metabolic organ that is essential for the regulation of whole-body glucose homeostasis. Dysfunctional adipose tissue hypertrophy with obesity is associated with glucose intolerance and insulin resistance underlying type 2 diabetes. Yes-associated protein 1 (YAP) is a transcription cofactor important in the Hippo signaling pathway. However, the role of YAP in adipose tissue and glucose homeostasis is unknown. To understand the role of YAP in vivo under metabolic stress conditions, we assessed how increased weight and insulin resistance impact YAP protein levels. We found that YAP protein levels increase in adipose tissue in humans with type 2 diabetes and mouse models. This suggests that YAP signaling may contribute to adipocyte dysfunction and insulin resistance under metabolic stress conditions. To further investigate the role of YAP in adipose tissue and glucose homeostasis in vivo, we developed adipose tissue-specific YAP knockout (YAP-KO) mice and placed them on either chow or high fat diet (HFD) for 12-14 weeks. On an HFD, adipose tissue YAP-KO mice show improved glucose tolerance compared to controls. Perigonadal fat pad weight was also decreased in knockout animals, with smaller adipocyte size. Adipose tissue fibrosis and gene expression associated with fibrosis was decreased in vivo and in 3T3-L1 cells treated with a YAP inhibitor. Together, these data indicate that YAP is increased in adipose tissue with weight gain and insulin resistance. Disruption of YAP in adipocytes improves glucose tolerance and adipose tissue fibrosis; suggesting that YAP plays an important role in adipose tissue and modulating glucose homeostasis under metabolic stress conditions.

Disclosure
D.Han: None. R.Aslam: None. T.Ojha: n/a. D.Yuen: Consultant; Fibrocor Therapeutics. C.T.Luk: None.

Funding
NSERC Discovery Grant (RGPIN-2018-05671) BBDC Gales Family Charitable Foundation Pilot and Feasibility GrantCanadian Institutes of Health Research Project Grant (PJT 168996)
24-LB: Guanfacine Normalizes Presynaptic Alpha-2 Adrenoceptors Enrichment and Ameliorates Neuropathic Pain in Type 1 Diabetic Rats
Authors: MUNAWAR NEHA , AL MADHOUN ASHRAF , NADER JOELLE , AL-ALI WALEED , MASOCHA WILLIAS , AL-MULLA FAHD , BITAR MILAD S. 
Published: May 2022

Diabetes is associated with several complications, including neuropathic pain, which is challenging to manage with currently available drugs. Descending noradrenergic neurons possess anti-nociceptive activity; however their involvement in diabetic neuropathic pain remains to be explored. To infer the regulatory role of this system, we examined in the pons, a part of the brainstem, lumbar nerves of the spinal cord and dorsal root ganglia of streptozotocin (STZ) -treated rats, a model for type 1 diabetes (T1D) , the localization (immunofluorescence) and the mRNA (qRT-PCR) and protein (Western blotting) expression of alpha-2A adrenoceptor (ADRA2A) . The data revealed that presynaptic SNAP-25 labeled ADRA2A in the central and peripheral nervous system of STZ diabetic rats were up-regulated both at the mRNA and protein levels. Interestingly, the levels of PSD-95 labeled postsynaptic neuronal ADRA2A remain unaltered as a function of diabetes. These biochemical abnormalities in the noradrenergic system of diabetic animals were associated with increased pain sensitivity as typified by the presence of hyperalgesia and cold/mechanical allodynia. The latter pain-related behaviors were assessed using Hargreaves apparatus, cold-plate and dynamic plantar aesthesiometer. Chronically administered guanfacine, a selective ADRA2A agonist, to diabetic animals downregulated the upregulation of neuronal presynaptic ADRA2A and ameliorated the hyperalgesia and the cold/mechanical allodynia in these animals. Together, these findings demonstrate that guanfacine may function as a potent analgesic and highlight ADRA2A, a key component of the descending neuronal autoinhibitory pathway as a potential therapeutic target in the treatment of diabetic neuropathic pain.

Disclosure
N. Munawar: None. A. Al madhoun: None. J. Nader: None. W. Al-ali: None. W. Masocha: None. F. Al-mulla: None. M. S. Bitar: None.

Funding
Grant YM05/19 from Kuwait University Research Sector.
247-LB: Pancreatic Elastase Regulates ß-Cell Viability by Impairing the Mechanosignaling Pathway
Authors: BASILE GIORGIO , VETERE AMEDEO , HU JIANG , IJADUOLA OLUWASEUN , DE JESUS DARIO F. , FUKUDA KAZUKI , ELTONY AMIRA M. , YUN ANDY , CHOUDHARY AMIT , WAGNER BRIDGET , KULKARNI ROHIT 
Published: May 2022

The discovery of SerpinB1 as an inducer of human β-cell proliferation pointed to acinar cell-derived proteases such as pancreatic elastase (PE) being involved in regulating β-cell viability. Indeed, we observed that expression of a major form of PE, namely CELA3B, was increased in acinar cells from donors with type 2 diabetes (T2D) compared to samples from non-diabetic (ND) donors. Surprisingly, CELA3B protein levels were higher in T2D vs. ND islets, although its gene expression levels were virtually undetectable. We confirmed that increasing doses of exogenous PE is able to decrease viability of EndoC-βH3 (βH3) cells in vitro. To evaluate the effects of PE inhibition on β-cell survival, we used telaprevir, an FDA-approved drug, that we identified as a potent PE blocker using a High-Throughput Screening assays with an IC50 of 15.6 nM to inhibit PE activity. Consistent with the mitogenic effects of SerpinB1 on β-cells, telaprevir also increased human β-cell viability in in vitro and in vivo systems. By performing phospho-antibody microarrays and single-cell RNA-Seq in human islets treated with vehicle or telaprevir, we identified the mechano-signaling pathway, which integrates the signals generated by the extracellular matrix (ECM) , as a candidate pathway linking PE inhibition to β-cell viability. We validated the results by treating EndoC-βH1 (βH1) cells lacking focal adhesion kinase (siFAK) with PE +/- telaprevir. We observed that PE impaired the mechano-signaling cascade, and these effects were reversed by telaprevir in control cells, but not in siFAK cells. Furthermore, using the novel approach of Brillouin microscopy we observed that PE increased the intracellular stiffness in βH1 cells, as a readout of cytoskeleton arrangements, whereas telaprevir blunted these effects. These data point to novel pathways underlying the acinar-β-cell crosstalk that could be harnessed for therapeutic approaches to resolve diabetes in patients.

Disclosure
G. Basile: None. B. Wagner: Research Support; Japan Tobacco Inc., Novo Nordisk Foundation, Ono Pharmaceutical Co., Ltd., Stock/Shareholder; Moderna, Inc., Pfizer Inc. R. Kulkarni: Advisory Panel; Biomea, Novo Nordisk, REDD Pharma, Consultant; Relay Therapeutics, Research Support; Inversago, SerPlus. A. Vetere: None. J. Hu: None. O. Ijaduola: None. D. F. De jesus: n/a. K. Fukuda: None. A. M. Eltony: None. A. Yun: None. A. Choudhary: None.

Funding
JDRF: 3-PDF-2020-935-A-NNIH: U01 DK123717NIH: UC4 DK116255
247-OR: Maternal Prepregnancy Obesity and Gestational Diabetes Are Associated with Epigenetic Gestational Aging in the Placenta
Authors: HUFF KATELYN , ROELL KYLE , BULKA CATHERINE , BOYLE KRISTEN E. , BRETON CARRIE , BURT AMBER , DABELEA DANA , KAHN LINDA , KARAGAS MARGARET R. , MARSIT CARMEN , NIEMIEC SIERRA , VOLK HEATHER , O'SHEA THOMAS M. , FRY REBECCA 
Published: May 2022

Pregnancy conditions like maternal pre-pregnancy obesity and gestational diabetes (GDM) have been linked with the development of placenta, a critical organ required for maternal-fetal interface during gestation, and can lead to adverse health outcomes in both mother and fetus. However, the molecular signatures associated with these conditions are not fully understood. Epigenetic clocks, which are based upon DNA methylation of age-related CpGs, are thought to better reflect biological age compared to chronological age and have been used in different tissues as potential biomarkers. In the present study, we hypothesized that altered placental epigenetic gestational aging is associated with pre-pregnancy obesity and GDM. We explored these relationships by leveraging data from three diverse cohorts in the Environmental influences on Child Health Outcomes (ECHO) program (overall N=830) . First, we investigated associations between maternal pre-pregnancy obesity and GDM and estimated placental epigenetic gestational age acceleration (eGAA) . Next, we examined associations between these two conditions and methylation of each CpG site within the robust placental clock (558 CpGs) . Significant associations were not observed between pre-pregnancy obesity and GDM and eGAA. Interestingly, pre-pregnancy obesity was associated with methylation levels at 28 placental clock CpGs (q&lt;0.1) . These sites were annotated to 17 unique genes that enrich processes like circadian entrainment and cellular response to glucagon stimulus (GNAS, GNG2) . Additionally, GDM was associated with methylation levels at 22 CpGs that mapped to 14 genes related to apoptosis (TNFRSF10B, TRADD) and neurological development (CACNA1G, ATXN1) . Overall, findings suggest that maternal pre-pregnancy obesity and GDM are associated with CpG methylation of the placental epigenetic clock, revealing potential markers vital for healthy pregnancy and fetal development.

Disclosure
K.Huff: None. C.Marsit: None. S.Niemiec: None. H.Volk: None. T.M.O'shea: None. R.Fry: None. K.Roell: None. C.Bulka: None. K.E.Boyle: None. C.Breton: None. A.Burt: None. D.Dabelea: None. L.Kahn: None. M.R.Karagas: None.

Funding
National Institutes of Health (UH3OD023348)
250-LB: Tead1—A Regulator of Adult ß-Cell Proliferation and Function
Authors: LEE JEONGKYUNG , LIU RUYA , LI FENG , NEGI VINNY , JAGANNATHAN RAJAGANAPATHI , HUISING MARK , MA KE , SHIH BEN , MOULIK MOUSUMI , YECHOOR VIJAY 
Published: May 2022

The transcriptional factor, Tead1, mediates the transcriptional output of the Hippo pathway. Tead1 activates the transcription of downstream genes including many cell cycle promoting genes that lead to cell proliferation in many tissues, but its role in β-cells is unknown. To test if Tead1 is required for β-cell proliferation and function, we deleted Tead1 in β-cell. Deletion of Tead1 in β-cells was confirmed in these β-cell Tead1 KO (β-Tead1-/-) , by a significant decrease in transcript and protein from whole islet lysates. The β-Tead1-/- mice had normal body weight but developed significantly higher fasting and fed glucose levels starting at 5 weeks of age and progressed to frank diabetes by 8 weeks, with fasting blood glucose &gt;300 mg/dl, accompanied by hypoinsulinemia, as compared to floxed control mice. 8wk β-Tead1-/- mice display severe glucose intolerance due to an abrogation of GSIS during GTT, with a loss of first phase insulin secretion. Pancreatic insulin content was decreased by ∼50%, due to a decrease in expression of mature β-cell genes including, Pdx1, Nkx6.1 and MafA, all of which, we show to be direct transcriptional targets of Tead1. Tead1 also binds to the proximal promoter at the Ink4a locus to regulate p16/p19, critical cell cycle inhibitors, and this loss of activation in Tead1-deficient β-cells leads to an enhanced entry into cell cycle. Furthermore, to test if Tead1 has a similar regulatory role in human β-cells, we deleted TEAD1 knockout in human iPS cell-derived β-cells using CRISPR/Cas9 system. Tead1 deficient iPS cell-derived β-cells displayed a significant decrease in vitro GSIS. Our results demonstrate Tead1 to be a transcriptional switch required for β-cells to maintain mature function and remain in proliferative quiescence. This novel regulatory pathway could be targeted for β-cell replacement therapy to achieve β-cell proliferation without a loss of mature function.

Disclosure
J. Lee: None. V. Yechoor: n/a. R. Liu: None. F. Li: None. V. Negi: None. R. Jagannathan: None. M. Huising: Consultant; Crinetics Pharmaceuticals, Inc., Research Support; Crinetics Pharmaceuticals, Inc. K. Ma: None. B. Shih: None. M. Moulik: None.

Funding
VA-MERIT Grant (I01BX002678)
251-LB: Tracking Beta-Cell Regeneration in Human Pancreatic Slices using Adenovirus Transduction
Authors: ALVAREZ-CUBELA SILVIA , KLEIN DAGMAR , ALVER CHARLES G. , MUHAMMAD FAHD QADIR MIRZA , MATEUS GONCALVES LUCIANA , QURESHI FARHAN , ALMACA JOANA , FRAKER CHRISTOPHER , CAICEDO ALEJANDRO , PUGLIESE ALBERTO , RICORDI CAMILLO , OLTRA ELISA , PUGLIESE ALBERTO , AGARWAL ASHUTOSH G. , PASTORI RICARDO , DOMÍNGUEZ-BENDALA JUAN 
Published: May 2022

Introduction: The study of pancreatic regeneration would benefit greatly from the design and validation of robust human-based models. Human Pancreatic Slices (HPSs) are thin organotypic sections of live pancreatic tissue. The sectioning method preserves the overall histological structure of the organ, maintaining the integrity of the extracellular matrix and the natural interaction between the endocrine and exocrine compartments, as well as the local neural, vascular and immune milieu. Conditions for the long-term culture of HPSs, recently reported by our team, have enabled the real-time analysis of beta-cell neogenesis using adenoviral (AV) co-transduction of a red-green reporter and an insulin tracer in human pancreatic slices. However, the ability of these cells to respond to glucose was not established at that time.
Methods: To determine whether new INS+ cells respond to glucose, we have designed an AV in which INS-dependent recombination leads to the expression of a blue marker (moxBFP) and a Calcium Imaging Reporter (gcAMP6s, green) whose intensity is proportional to glucose-dependent INS secretion. This allowed us to monitor glucose-stimulated calcium influx in the newly created cells. To track the generation of new beta cells with an even higher degree of resolution we subsequently placed the transduced slices in a custom-made microfluid chip.
Conclusions: This setting further allows for longitudinal functional analyses in precise and highly controlled experimental conditions. Our ability to study regeneration in a clinically meaningful model represents a groundbreaking advance that may fast-track the screening and preclinical development of therapeutic agents.

Disclosure
S. Alvarez-cubela: n/a. A. Pugliese: Advisory Panel; Provention Bio, Inc., Consultant; Quell Tx. C. Ricordi: Advisory Panel; Vertex Pharmaceuticals Incorporated. E. Oltra: None. A. Pugliese: Advisory Panel; Provention Bio, Inc., Consultant; Quell Tx. A. G. Agarwal: None. R. Pastori: None. J. Domínguez-bendala: None. D. Klein: None. C. G. Alver: None. M. Qadir: None. L. Mateus goncalves: None. F. Qureshi: None. J. Almaca: None. C. Fraker: None. A. Caicedo: None.

Funding
National Institutes of Health (RO1DK130846UO1DK12393)
252-OR: Maternal Obesity and Diabetes during Pregnancy and Quantitative Checklist for Autism in Toddlers (QCHAT) Scores in Offspring
Authors: CARTER SARAH A. , LIN JANE C. , CHOW TING , MARTINEZ MAYRA P. , ALVES JASMIN M. , FELDMAN R. KLARA , QIU CHUNYUAN , PAGE KATHLEEN A. , XIANG ANNY 
Published: May 2022

In utero exposure to maternal obesity and diabetes has been associated with risk of autism spectrum disorders (ASD) in children. As early identification and support improves long-term outcomes in children with ASD, a 10-point Quantitative Checklist for Autism in Toddlers (QCHAT) screening questionnaire was developed and validated for clinical screening between 18-24 months at well-child visits. This study assessed associations between maternal metabolic exposures and offspring performance on early QCHAT screening. Data included 65699 singletons born in a single healthcare system, with QCHAT administered toddler well-child visits. Demographic and social data, maternal pregnancy obesity (BMI≥30 kg/m2) , pre-existing type 1/type 2 (T1D/T2D) , and gestational diabetes (GDM) status, and child’s QCHAT score were extracted from electronic medical records. Negative binomial regression was used to assess associations between maternal obesity and diabetes and QCHAT scores as a continuous measure, adjusting for covariates. Analyses included and excluded children with ASD diagnoses and were run with and without mutual adjustment for each metabolic condition. In this cohort, 18923 (28.8%) of children were exposed to maternal obesity, 2564 (3.9%) to T1/T2, 1946 (3.0%) to GDM diagnosed at ≤26 weeks, and 3290 (5.0%) GDM diagnosed &gt;26 weeks. Of the total cohort, 1622 (2.5%) were later diagnosed with ASD. Maternal obesity (Incidence Rate Ratio: 1.12, 95% CI: 1.09,1.14) , T1D/T2D (1.13, 1.07-1.20) , and GDM ≤26 weeks (1.12, 1.05-1.19) were associated with one-unit increases in QCHAT scores. No associations were reported for GDM &gt;26 weeks. Similar patterns were observed after excluding children with ASD diagnoses. Mutual adjustment did not change results. These data suggest maternal metabolic conditions during pregnancy are associated with increased social and communication impairment on early autism screening tools, even in typically developing children.

Disclosure
S.A.Carter: None. J.C.Lin: None. T.Chow: None. M.P.Martinez: None. J.M.Alves: None. R.Feldman: None. C.Qiu: None. K.A.Page: None. A.Xiang: None.
253-LB: Ethnic Differences in Pancreatic Hormone Secretion in Health and T2D
Authors: DOLIBA NICOLAI M. , ROMAN JEFFREY , ROZO ANDREA V. , QIN WEI , LIU CHENGYANG , NAJI ALI , RICKELS MICHAEL R. , ATKINSON MARK A. , POWERS ALVIN C. , BRISSOVA MARCELA , KAESTNER KLAUS H. , STOFFERS DORIS A. 
Published: May 2022

Purpose: We tested the hypothesis that a critical biologic determinant of T2D disparities is based on ethnicity-related differences in pancreatic islet function.
Methods: Utilizing donor tissues from the NIDDK-funded Human Pancreas Analysis Program (HPAP) , we examined both insulin and glucagon secretion from nondiabetic and diabetic Caucasian and African American donor islets. Initially, a physiological amino acid mixture was used to stimulate glucagon secretion, followed by low and high glucose to stimulate insulin secretion and suppress glucagon secretion. During the high glucose step, IBMX (0.1 mM) is added to potentiate secretion of both hormones. Insulin and glucagon concentration in perifusates and islet extracts was measured by radioimmunoassay or ELISA. Statistical comparisons were drawn by repeated measures ANOVA.
Results: Comparing nondiabetic islets, we observed overlapping insulin secretion profiles between both ethnicities; however, glucagon secretion was distinctly greater in both nondiabetic African American (+111%) islets compared to nondiabetic Caucasian islets, under all interventions. In T2D, the reduction in insulin secretion was significantly greater in African American T2D donors (high glucose -76%; IBMX -76%) compared to Caucasian T2D donors (high glucose -47%; IBMX -50%) . More strikingly, glucagon secretion in T2D donors was markedly different based on ethnicity. Whereas Caucasian T2D islets exhibited a similar baseline of glucagon secretion compared to Caucasian controls, there was a marked reduction of glucagon secretion overall in African American T2D donors (-70%) compared to their corresponding controls. The largest detected difference was in IBMX-potentiation of glucagon secretion, which is higher in Caucasian T2D donors (+103%) but significantly decreased in African American T2D donors (-76%) . Preliminary analysis of hormone content in a subset of the donor islet preparations revealed no differences according to ethnicity.
Conclusions: We propose that ethnicity-related differences in nondiabetic islet function may contribute to the enhanced risk of T2D.

Disclosure
N. M. Doliba: None. M. Brissova: None. K. H. Kaestner: None. D. A. Stoffers: Other Relationship; Biomarin, Correlagen. J. Roman: None. A. V. Rozo: None. W. Qin: None. C. Liu: None. A. Naji: None. M. R. Rickels: Advisory Panel; Sernova, Corp., Vertex Pharmaceuticals Incorporated, Zealand Pharma A/S, Consultant; L-Nutra Inc. M. A. Atkinson: None. A. C. Powers: None.

Funding
This work is supported by 3UC4-112217-01S1 and U01-DK123594-02, UC4-DK-112217, UC4-DK-112232, and U01-DK-123716.
305-OR: Quiescence Abrogates Stress-Induced Senescence-Associated Secretion Phenotype in Adipose-Derived Stem Cells and May Enhance Their Therapeutic Effects on Diabetic Complications
Authors: FUNAKI MAKOTO , HATA AKIKO , YOSHIDA MITSUTERU , MIYAMOTO NAOKI , IKUTA TAKAHISA , UEMURA YUTA , AKIZUKI MINATO , TANGOKU AKIRA , YUMOTO HIROMICHI , OGAWA HIROHISA , OYA TAKESHI 
Published: May 2022

The anti-inflammatory functions of adipose-derived stem cells (ADSC) are receiving much attention as potential treatment for diabetic complications, although it is not yet established. Cellular stresses render ADSC prematurely senescent, in which senescent ADSC propagate instead of suppressing inflammation in neighboring cells and tissues through their secretory profile, which is called senescence-associated secretion phenotype (SASP) . In diabetes, ADSC exhibit SASP, which decreases the feasibility of ADSC-based therapies. We previously reported that ADSC become quiescent when cultured on a substrate whose stiffness mimics that of adipose tissue. Here, we investigated the effect of quiescence on high glucose-induced SASP in ADSC. When cultured in three-dimensional (3D) gels, ADSC became quiescent and showed upregulated indolemine 2,3-dioxygene in response to tumor necrosis factor α and interferon γ stimulation to mimic inflammation ex vivo. High glucose treatment upregulated MCP-1 in ADSC cultured on plastic tissue culture plates, and their conditioned medium decreased the viability of HUVEC, an endothelial cell line. However, even under high glucose conditions, transferring ADSC from plastic to 3D gels attenuated MCP-1 expression, and their conditioned medium restored the viability of HUVEC. Transplantation of gels containing ADSC into mice resulted in vascular formation in those gels, and enhanced wound healing was suggested. These results open up the possibility that quiescence will eliminate SASP in ADSC even under hyperglycemia and promote the ability of ADSC to have therapeutic effects on diabetic complications.

Disclosure
M.Funaki: Board Member; Mechanogenic, Inc. H.Ogawa: None. T.Oya: None. A.Hata: None. M.Yoshida: None. N.Miyamoto: None. T.Ikuta: None. Y.Uemura: None. M.Akizuki: None. A.Tangoku: None. H.Yumoto: None.