Diabetes最新文献

Loss of β-cell KATP reduces Ca2+ sensitivity of insulin secretion and Trpm5 expression

IF 7.7 1区医学

Diabetes Pub Date : 2024-12-12 DOI: 10.2337/db24-0650

Nathaniel W. York, Zihan Yan, Anna B. Osipovich, Abbie Tate, Sumit Patel, David W. Piston, Mark A. Magnuson, Maria S. Remedi, Colin G. Nichols

{"title":"Loss of β-cell KATP reduces Ca2+ sensitivity of insulin secretion and Trpm5 expression","authors":"Nathaniel W. York, Zihan Yan, Anna B. Osipovich, Abbie Tate, Sumit Patel, David W. Piston, Mark A. Magnuson, Maria S. Remedi, Colin G. Nichols","doi":"10.2337/db24-0650","DOIUrl":"https://doi.org/10.2337/db24-0650","url":null,"abstract":"Loss-of-function (LOF) mutations in KATP channels cause hyperexcitability and insulin hypersecretion, resulting in congenital hyperinsulinism (CHI). Paradoxically, despite the initial insulin hypersecretion, many CHI cases, as well as KATP knockout (KO) animals, eventually ‘crossover’ to undersecretion and even diabetes. Here we confirm that Sur1 KO islets exhibit higher intracellular [Ca2+] ([Ca2+]i) at all [glucose], but show decreased glucose-stimulated insulin secretion. However, when [Ca2+]i is artificially elevated by increasing extracellular [Ca2+], insulin secretion from Sur1 KO islets increases to the same levels as WT islets. This indicates that a right-shift in [Ca2+]i-dependence of insulin secretion, rather than loss of insulin content or intrinsic secretability, is the primary cause for the crossover. Chronic pharmacological inhibition of KATP channel activity by slow release of glibenclamide in pellet-implanted mice causes a very similar ‘crossover’ to glucose intolerance and impaired insulin secretion to that seen in Sur1 KO animals. Whole islet and single cell transcriptomic analysis reveal markedly reduced Trpm5 in both conditions. Glibenclamide pellet-implanted Trpm5 KO mice also exhibited significant glucose intolerance. However, this was not as severe as in WT animals, which suggests that decreased expression of Trpm5 may play a small role in the disruption of insulin secretion with KATP loss.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"11 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142815501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Adipose tissue biology and effect of weight loss in women with lipedema

IF 7.7 1区医学

Diabetes Pub Date : 2024-12-09 DOI: 10.2337/db24-0890

Vincenza Cifarelli, Gordon I. Smith, Silvia Gonzalez-Nieves, Dmitri Samovski, Hector H. Palacios, Jun Yoshino, Richard I. Stein, Anja Fuchs, Thomas F. Wright, Samuel Klein

{"title":"Adipose tissue biology and effect of weight loss in women with lipedema","authors":"Vincenza Cifarelli, Gordon I. Smith, Silvia Gonzalez-Nieves, Dmitri Samovski, Hector H. Palacios, Jun Yoshino, Richard I. Stein, Anja Fuchs, Thomas F. Wright, Samuel Klein","doi":"10.2337/db24-0890","DOIUrl":"https://doi.org/10.2337/db24-0890","url":null,"abstract":"Lipedema is a lipodystrophic disease that is typically characterized by a marked increase in lower-body subcutaneous adipose tissue that is purported to have increased inflammation and fibrosis, impaired microvascular/lymphatic circulation and to be resistant to reduction by weight loss therapy. However, these outcomes have not been adequately studied. We evaluated body composition, insulin sensitivity, metabolic health and adipose tissue biology in women with obesity and lipedema (Obese-LIP) before and after moderate (~9%) diet-induced weight loss. At baseline, people with Obese-LIP had ~23% greater leg fat mass, ~11% lower android-to-gynoid ratio and ~48% greater insulin sensitivity (all P&lt;0.05) than women matched on age, BMI and whole-body adiposity. In Obese-LIP, macrophage content and expression of genes involved in inflammation and fibrosis were greater, whereas lymph/angiogenesis-related genes were lower in thigh than abdominal subcutaneous adipose tissue. Weight loss improved insulin sensitivity and decreased total fat mass, with similar relative reductions in abdominal and leg fat masses, but without changes in markers of inflammation and fibrosis. These results demonstrate that affected adipose tissue in women with lipedema is characterized by increased inflammation and fibrogenesis, and alterations in lymphatic and vascular biology. Moderate diet-induced weight loss improves metabolic function and decreases lower-body adipose tissue mass.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"37 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142797021","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Gain of function NOTCH3 variants cause familial partial lipodystrophy due to activation of senescence pathways

IF 7.7 1区医学

Diabetes Pub Date : 2024-12-09 DOI: 10.2337/db24-0624

Abhimanyu Garg, Chao Xing, Anil K. Agarwal, Aundrea K. Westfall, Diana R. Tomchick, Xunzhi Zhang, Michelle Xing, Rebecca J. Brown

{"title":"Gain of function NOTCH3 variants cause familial partial lipodystrophy due to activation of senescence pathways","authors":"Abhimanyu Garg, Chao Xing, Anil K. Agarwal, Aundrea K. Westfall, Diana R. Tomchick, Xunzhi Zhang, Michelle Xing, Rebecca J. Brown","doi":"10.2337/db24-0624","DOIUrl":"https://doi.org/10.2337/db24-0624","url":null,"abstract":"Despite elucidation of the molecular genetic basis of several lipodystrophy syndromes, molecular defects in some ultra-rare subtypes of familial lipodystrophies remain unidentified. We analyzed whole exome sequencing (WES) data of four affected and two unaffected females from an undiagnosed autosomal dominant familial partial lipodystrophy (FPL) pedigree and identified only one novel heterozygous variant, p.Ala1603Tyr in NOTCH3 meeting the filtering criteria. Further analysis of WES data of 222 unexplained FPL patients identified two unrelated FPL patients with novel heterozygous, p.Cys1600Tyr and p.Gln1552Pro, NOTCH3 variants. All variants were clustered in the heterodimerization domain of the negative regulatory region of NOTCH3. RNA-Seq and proteomics analysis of skin fibroblasts revealed significantly higher RNA and protein expression of NOTCH3 and activation of widespread senescence pathways in the FPL patients versus controls. NOTCH3 is highly expressed in adipose tissue and plays many crucial roles in developmental patterning, cell fate decisions, regulation of cell survival and proliferation. We conclude that gain-of-function missense variants in the negative regulatory region of NOTCH3 cause a novel subtype of FPL by activation of senescence pathways. This novel variety of FPL should be considered for non-obese patients with early- or childhood-onset diabetes mellitus.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"7 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-12-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142797020","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

β-Cell Secretory Capacity Predicts Metabolic Outcomes over 6 Years following Human Islet Transplantation

IF 7.7 1区医学

Diabetes Pub Date : 2024-12-04 DOI: 10.2337/db24-0729

Anneliese J. Flatt, Austin M. Matus, Robert J. Gallop, Eileen Markmann, Cornelia Dalton-Bakes, Amy J. Peleckis, Chengyang Liu, Ali Naji, Michael R. Rickels

{"title":"β-Cell Secretory Capacity Predicts Metabolic Outcomes over 6 Years following Human Islet Transplantation","authors":"Anneliese J. Flatt, Austin M. Matus, Robert J. Gallop, Eileen Markmann, Cornelia Dalton-Bakes, Amy J. Peleckis, Chengyang Liu, Ali Naji, Michael R. Rickels","doi":"10.2337/db24-0729","DOIUrl":"https://doi.org/10.2337/db24-0729","url":null,"abstract":"Transplanted islet functional β-cell mass is measured by the β-cell secretory capacity derived from the acute insulin response to glucose-potentiated arginine (AIRpot), however, data are limited beyond one-year post-transplant for individuals with type 1 diabetes. We evaluated changes in β-cell secretory capacity in a single-center longitudinal analysis and examined relationships with measures of islet cell hormone metabolism and clinical measures of graft function (mixed-meal tolerance test [MMTT] C-peptide, BETA-2 score, and continuous glucose monitoring [CGM]). Eleven individuals received purified human pancreatic islets over one or two intra-portal infusions to achieve insulin-independence and were followed over a median (IQR) 6 (5-7) years. β-cell secretory capacity remained stable over 3-years before declining. Fasting glucagon and proinsulin secretory ratios under glucose-potentiation were inversely correlated with AIRpot. A functional β-cell mass of 40% normal predicted insulin-independence and was strongly predicted by MMTT C-peptide-to-glucose and BETA-2 score. A functional β-cell mass of &gt;20% predicted excellent glycemic outcomes including ≤1% time &lt;60 mg/dL, ≤2% time &gt;180 mg/dL and ≥90% time-inrange 70-180 mg/dL. β-cell replacement approaches should target a functional β-cell mass &gt;40% to provide sufficient islet reserve for sustained insulin-independence. MMTT C-peptide-to-glucose and BETA-2 score can inform changes in functional β-cell mass in the clinical setting.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"4 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142776688","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Plasma proteomic signatures of adiposity are associated with cardiovascular risk factors and type 2 diabetes risk in a multi-ethnic Asian population.

IF 7.7 1区医学

Diabetes Pub Date : 2024-12-02 DOI: 10.2337/db24-0184

Charlie G.Y. Lim, Bige Ozkan, Yujian Liang, Jingsha Chen, Jiali Yao, Nang Ei Ei Khaing, Mary R. Rooney, Chiadi E. Ndumele, E Shyong Tai, Josef Coresh, Xueling Sim, Rob M. van Dam

{"title":"Plasma proteomic signatures of adiposity are associated with cardiovascular risk factors and type 2 diabetes risk in a multi-ethnic Asian population.","authors":"Charlie G.Y. Lim, Bige Ozkan, Yujian Liang, Jingsha Chen, Jiali Yao, Nang Ei Ei Khaing, Mary R. Rooney, Chiadi E. Ndumele, E Shyong Tai, Josef Coresh, Xueling Sim, Rob M. van Dam","doi":"10.2337/db24-0184","DOIUrl":"https://doi.org/10.2337/db24-0184","url":null,"abstract":"The biomarkers connecting obesity and cardiometabolic diseases are not fully understood. We aimed to (i) evaluate the associations between body mass index (BMI), waist circumference (WC), and ∼5,000 plasma proteins (SomaScan v4), (ii) identify protein signatures of BMI and WC, and (iii) evaluate the associations between the protein signatures and cardiometabolic health including metabolically unhealthy obesity and type 2 diabetes incidence in the Singapore Multi-Ethnic Cohort (MEC1). Among 410 BMI-associated and 385 WC-associated proteins, we identified protein signatures of BMI and WC and validated them in an independent dataset across two timepoints and externally in the Atherosclerosis Risk in Communities (ARIC) study. The BMI- and WC-protein signatures were highly correlated with total and visceral body fat, respectively. Furthermore, the protein signatures were significantly associated with cardiometabolic risk factors and metabolically unhealthy obesity. In prospective analyses, the protein signatures were strongly associated with type 2 diabetes risk in MEC1 (odds ratio per SD increment in WC-protein signature = 2.84, 95% CI 2.47 to 3.25) and ARIC (hazard ratio = 1.98, 95% CI 1.88 to 2.08). Our protein signatures have potential uses for the monitoring of metabolically unhealthy obesity.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"46 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142760367","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Blocking adipocyte YY1 decouples thermogenesis from beneficial metabolism by promoting spermidine production

IF 7.7 1区医学

Diabetes Pub Date : 2024-12-02 DOI: 10.2337/db24-0501

Chen Qiu, Yu Lu, Suyang Wu, Wenli Guo, Jiahao Ni, Jiyuan Song, Zichao Liu, Xiaoai Chang, Kai Wang, Peng Sun, Qian Zhang, Shufang Yang, Kai Li

{"title":"Blocking adipocyte YY1 decouples thermogenesis from beneficial metabolism by promoting spermidine production","authors":"Chen Qiu, Yu Lu, Suyang Wu, Wenli Guo, Jiahao Ni, Jiyuan Song, Zichao Liu, Xiaoai Chang, Kai Wang, Peng Sun, Qian Zhang, Shufang Yang, Kai Li","doi":"10.2337/db24-0501","DOIUrl":"https://doi.org/10.2337/db24-0501","url":null,"abstract":"The accumulation of mitochondria in thermogenic adipose tissue (i.e., brown and beige fat) increases energy expenditure, which can aid in alleviating obesity and metabolic disorders. However, recent studies have shown that knocking out key proteins required to maintain mitochondrial function inhibits the energy expenditure in thermogenic fat, and yet the knockout mice are unexpectedly protected from developing obesity or metabolic disorders when fed a high-fat diet (HFD). In the present study, non-biased sequencing-based screening revealed the importance of YY1 in the transcription of electron transport chain genes and the enhancement of mitochondrial function in thermogenic adipose tissue. Specifically, adipocyte YY1 null (YAKO) mice showed lower energy expenditure and were intolerant to cold stress. Interestingly, YAKO mice showed alleviation of HFD-induced metabolic disorders, which can be attributed to a suppression of adipose tissue inflammation. Metabolomic analysis revealed that blocking YY1 directed glucose metabolism toward lactate, enhanced the uptake of glutamine, and promoted the production of anti-inflammatory spermidine. Conversely, blocking spermidine production in YAKO mice reversed their resistance to HFD-induced disorders. Thus, although blocking adipocyte YY1 impairs the thermogenesis, it promotes spermidine production and alleviates adipose tissue inflammation, therefore leads to an uncoupling of adipose tissue energy expenditure from HFD-induced metabolic disorders.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"82 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142760505","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Resolving spatiotemporal electrical signaling within the islet via CMOS microelectrode arrays 通过 CMOS 微电极阵列解析胰岛内的时空电信号

IF 7.7 1区医学

Diabetes Pub Date : 2024-11-25 DOI: 10.2337/db23-0870

Anne Gresch, Jana Osthues, Jan D. Hüwel, Jennifer K. Briggs, Tim Berger, Ruben Koch, Thomas Deickert, Christian Beecks, Richard K.P. Benninger, Martina Düfera

{"title":"Resolving spatiotemporal electrical signaling within the islet via CMOS microelectrode arrays","authors":"Anne Gresch, Jana Osthues, Jan D. Hüwel, Jennifer K. Briggs, Tim Berger, Ruben Koch, Thomas Deickert, Christian Beecks, Richard K.P. Benninger, Martina Düfera","doi":"10.2337/db23-0870","DOIUrl":"https://doi.org/10.2337/db23-0870","url":null,"abstract":"Glucose-stimulated beta-cells exhibit synchronized calcium dynamics across the islet that recruit beta-cells to enhance insulin secretion. Compared to calcium dynamics, the formation and cell-to-cell propagation of electrical signals within the islet are poorly characterized. To determine factors that influence the propagation of electrical activity across the islet underlying calcium oscillations and beta-cell synchronization, we used high-resolution CMOS multielectrode arrays (MEA) to measure voltage changes associated with the membrane potential of individual cells within intact C57BL6 mouse islets. We measured fast (milliseconds, spikes) and slow (seconds, waves) voltage dynamics. Single spike activity and wave signal velocity were both glucose-dependent, but only spike activity was influenced by NMDA receptor activation or inhibition. A repeated glucose stimulus revealed a highly responsive subset of cells in terms of spike activity. When islets were pretreated for 72 hours with glucolipotoxic medium, the wave velocity was significantly reduced. Network analysis confirmed that in response to glucolipotoxicity the synchrony of islet cells was affected due to slower propagating electrical waves and not due to altered spike activity. In summary, this approach provided novel insight regarding the propagation of electrical activity and the disruption of cell-to-cell communication due to excessive stimulation.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"6 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-11-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142712608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Pre-clinical development of a tolerogenic peptide from glutamate decarboxylase as a candidate for antigen-specific immunotherapy in type 1 diabetes 作为 1 型糖尿病抗原特异性免疫疗法候选药物的谷氨酸脱羧酶耐受肽的临床前开发

IF 7.7 1区医学

Diabetes Pub Date : 2024-11-21 DOI: 10.2337/db23-0996

Sky T. H. Ng, Michael J. Price, Naomi Richardson, Maher Nawaf, Alastair Copland, Heather B. Streeter, Parth Narendran, David C. Wraith

{"title":"Pre-clinical development of a tolerogenic peptide from glutamate decarboxylase as a candidate for antigen-specific immunotherapy in type 1 diabetes","authors":"Sky T. H. Ng, Michael J. Price, Naomi Richardson, Maher Nawaf, Alastair Copland, Heather B. Streeter, Parth Narendran, David C. Wraith","doi":"10.2337/db23-0996","DOIUrl":"https://doi.org/10.2337/db23-0996","url":null,"abstract":"Dysregulation and loss of immune tolerance towards pancreatic β-cell autoantigens are features of type 1 diabetes (T1D). Until recently, life-long insulin injection was the only approved treatment for T1D, and this does not address the underlying disease pathology. Antigen-specific immunotherapy (ASI) seeks to restore tolerance and holds potential as a new therapeutic strategy for treating autoimmune diseases with well characterised antigens. Peptide ASI using processing independent CD4+ T-cell epitopes (PIPs) shows promising results in several autoimmune diseases. Here we successfully applied the principles of PIP design to the T1D autoantigen glutamate decarboxylase 65 (GAD65). Peptides spanning GAD65 predicted to be pan-HLA-DR binding were selected. Peptide P10 displayed enriched responses in peripheral blood mononuclear cells from people with T1D. The minimal epitope of the P10 peptide was fine mapped using T-cell hybridomas generated from HLA-DRB1*04:01 transgenic mice. This minimal epitope, P10Sol, was demonstrated to induce tolerance to the parent peptide in HLA-DRB1*04:01 transgenic mice using a novel activation-induced marker assay. Finally, we show that GAD65 P10Sol PIP is recognised by CD4+ T-cells from people with T1D who possess a range of HLA-DR alleles and can, therefore, be defined as a pan-DR binding peptide with therapeutic potential.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"66 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142684150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The IsletTester mouse: an immunodeficient model with stable hyperglycemia for the study of human islets IsletTester小鼠：用于研究人类胰岛的具有稳定高血糖的免疫缺陷模型

IF 7.7 1区医学

Diabetes Pub Date : 2024-11-21 DOI: 10.2337/db23-0887

Eric L. Waite, Mark Tigue, Ming Yu, Deeksha Lahori, Kai Kelly, Catherine Lee May, Ali Naji, Jeffrey Roman, Nicolai Doliba, Dana Avrahami, Kim-Vy Nguyen-Ngoc, Maike Sander, Benjamin Glaser, Klaus H. Kaestner

{"title":"The IsletTester mouse: an immunodeficient model with stable hyperglycemia for the study of human islets","authors":"Eric L. Waite, Mark Tigue, Ming Yu, Deeksha Lahori, Kai Kelly, Catherine Lee May, Ali Naji, Jeffrey Roman, Nicolai Doliba, Dana Avrahami, Kim-Vy Nguyen-Ngoc, Maike Sander, Benjamin Glaser, Klaus H. Kaestner","doi":"10.2337/db23-0887","DOIUrl":"https://doi.org/10.2337/db23-0887","url":null,"abstract":"The gold standard for assessing the function of human islets or β-like cells derived from stem cells involves their engraftment under the kidney capsule of hyperglycemic, immunodeficient mice. Current models, such as Streptozotocin (STZ) treatment of severely immunodeficient mice or the NRG-Akita strain are limited due to unstable and variable hyperglycemia and/or high morbidity of these models. To address these limitations, we developed the IsletTester mouse via CRISPR-Cas9 mediated gene editing of glucokinase (Gck), the glucose sensor of the β-cells, directly in NSG zygotes. IsletTester mice are heterozygous for an Arg345-&gt;stop mutation in Gck and present with stable random hyperglycemia (∼250mg/dl; ∼14 mM), normal life span and fertility. We demonstrate the utility of this model through functional engraftment of both human islets and hESC-derived β-like cells. The IsletTester mouse will enable the study of human islet biology over time and under different physiological conditions and can provide a useful preclinical platform to determine the functionality of stem cell-derived islet products.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"7 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142684215","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Tracking insulin- and glucagon-expressing cells in vitro and in vivo using a double reporter human embryonic stem cell line 利用双报告基因人类胚胎干细胞系跟踪体外和体内的胰岛素和胰高血糖素表达细胞

IF 7.7 1区医学

Diabetes Pub Date : 2024-11-19 DOI: 10.2337/db24-0756

Samantha Mar, Ekaterina Filatov, Shugo Sasaki, Majid Mojibian, Dahai Zhang, Angela Yang, Cuilan Nian, Francis C. Lynn

{"title":"Tracking insulin- and glucagon-expressing cells in vitro and in vivo using a double reporter human embryonic stem cell line","authors":"Samantha Mar, Ekaterina Filatov, Shugo Sasaki, Majid Mojibian, Dahai Zhang, Angela Yang, Cuilan Nian, Francis C. Lynn","doi":"10.2337/db24-0756","DOIUrl":"https://doi.org/10.2337/db24-0756","url":null,"abstract":"Human embryonic stem cell (hESC)-derived pancreatic alpha and beta cells can be used to develop cell replacement therapies to treat diabetes. However, recent published differentiation protocols yield varying amounts of alpha and beta cells amidst heterogeneous cell populations. To visualize and isolate hESC-derived alpha and beta cells, we generated a GLUCAGON-2AmScarlet and INSULIN-2A-EGFP dual fluorescent reporter (INSEGFPGCGmScarlet) hESC line using CRISPR/Cas9. We established robust expression of EGFP and mScarlet fluorescent proteins in insulin- and glucagon-expressing cells respectively without compromising the differentiation or function of these cells. We also showed the insulin- and glucagon-expressing bihormonal population at the maturing endocrine cell stage (Stage 6) of our pancreatic islet differentiation lose insulin expression over time, while maintaining an alpha-like expression profile, suggesting these bihormonal cells are cell autonomously fated to become alpha-like cells. We also demonstrated this cell line can be used to monitor hESC-derived insulin- and glucagonexpressing cells, and hESC-derived islet morphology in vivo by transplanting them into the anterior chamber of the eye in mice. Together, the INSEGFPGCGmScarlet hESC line provides an efficient strategy for tracking populations of hESC-derived beta- and alpha-like cells.","PeriodicalId":11376,"journal":{"name":"Diabetes","volume":"3 1","pages":""},"PeriodicalIF":7.7,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142673899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0