Identification of unique cell type responses in pancreatic islets to stress

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: July 2, 2024

Abstract Diabetes involves the death or dysfunction of pancreatic β-cells. Analysis bulk sequencing from human samples and studies using in vitro vivo models suggest that endoplasmic reticulum inflammatory signaling play an important role diabetes progression. To better characterize cell type-specific stress response, we perform multiplexed single-cell RNA to define transcriptional signature primary islet cells exposed stress. Through comprehensive pair-wise analysis responses across endocrine exocrine types, changes gene expression for each type under different diabetes-associated stressors. We find β-, α-, ductal have greatest response. utilize stem cell-derived islets study health through candidate CIB1 , which was upregulated islets. Our findings provide insights into establish a resource identify targets therapeutics.

Language: Английский

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SPOCK2 controls the proliferation and function of immature pancreatic β-cells through MMP2

Experimental & Molecular Medicine, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Human pluripotent stem cell-derived β-cells (SC-β-cells) represent an alternative cell source for transplantation in diabetic patients. Although mitogens could theory be used to expand β-cells, adult very rarely replicate. In contrast, newly formed including SC-β-cells, display higher proliferative capacity and distinct transcriptional functional profiles. Through bidirectional expression modulation single-cell RNA-seq, we identified SPOCK2, ECM protein, as inhibitor of immature β-cell proliferation. lacking SPOCK2 presented elevated MMP2 activity, leading β-integrin-FAK-c-JUN pathway activation. Treatment with the protein resulted pronounced short- long-term SC-β-cell expansion, significantly increasing glucose-stimulated insulin secretion vitro vivo. These findings suggest that mediates fetal proliferation maturation. summary, a molecular mechanism specifically regulates function, highlighting unique signaling milieu SC-β-cells promise robust derivation fully cells transplantation.

Language: Английский

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Clinically compliant enrichment of human pluripotent stem cell–derived islets

Science Translational Medicine, Journal Year: 2025, Volume and Issue: 17(792)

Published: April 2, 2025

Human pluripotent stem cell–derived islet (SC-islet) transplantation is a promising β cell replacement therapy for patients with type 1 diabetes, offering potential unlimited supply. Yet, the heterogeneity of final product containing non–target types has relevant implications SC-islet function, transplant volume, and safety. Here, we present clinically compliant, full three-dimensional differentiation protocol that includes purification step endocrine cell–rich clusters, relying on principle isopycnic centrifugation (density gradient separation). Enriched SC-islets displayed signs functionality in vitro vivo. In contrast antibody-based single-cell sorting approaches, this method does not destroy cytoarchitecture associated alterations function loss. Furthermore, it fast, easily scalable to large volumes, can be applied during manufacturing. This may also contribute generation improved cell-based therapies regenerative medicine purposes beyond field.

Language: Английский

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Recent progress in modeling and treating diabetes using stem cell-derived islets

Stem Cells Translational Medicine, Journal Year: 2024, Volume and Issue: 13(10), P. 949 - 958

Published: Aug. 19, 2024

Abstract Stem cell-derived islets (SC-islets) offer the potential to be an unlimited source of cells for disease modeling and treatment diabetes. SC-islets can genetically modified, treated with chemical compounds, or differentiated from patient derived stem model These models provide insights into pathogenesis vulnerabilities that may targeted treatment. themselves are also being investigated as a cell therapy However, transplantation process is imperfect; side effects immunosuppressant use have reduced SC-islet therapeutic potential. Alternative methods this include encapsulation, immunomodulating molecules, genetic modification SC-islets. This review covers recent advances using understand different diabetes pathologies therapy.

Language: Английский

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Untangling the genetics of beta cell dysfunction and death in type 1 diabetes

Molecular Metabolism, Journal Year: 2024, Volume and Issue: 86, P. 101973 - 101973

Published: June 22, 2024

Type 1 diabetes (T1D) is a complex multi-system disease which arises from both environmental and genetic factors, resulting in the destruction of insulin-producing pancreatic beta cells. Over past two decades, human studies have provided new insight into etiology T1D, including an appreciation for role cells their own demise. Here, we outline models supported by data cell dysfunction death T1D. We highlight importance strong evidence linking T1D associations to bona fide candidate genes mechanistic therapeutic consideration. To guide rigorous interpretation associations, describe molecular profiling approaches, genomic resources, that may be used construct variant-to-gene links investigate profile advances understanding causes at individual risk loci. introduce prediction discuss how they can address heterogeneity. Finally, present areas where investment will critical future use genetics open questions develop treatment prevention strategies

Language: Английский

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Controlling human stem cell-derived islet composition using magnetic sorting

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 21, 2024

Stem cell-derived islets (SC-islets) consists of multiple hormone-producing cell types and offer a promising therapeutic avenue for treating type 1 diabetes (T1D). Currently, the composition generated within these SC-islets currently cannot be controlled via soluble factors during this differentiation process consist off-target types. In study, we devised magnetic-activated sorting (MACS) protocol to enrich CD49a, marker associated with functional insulin-producing β cells. were from human pluripotent stem cells (hPSCs) using an adherent then sorted aggregated into islet-like clusters produce CD49a-enriched, CD49a-depleted, unsorted SC-islets. Single-cell RNA sequencing (scRNA-seq) immunostaining revealed that CD49a-enriched had higher proportions improved transcriptional identity compared other Functional assays demonstrated exhibited enhanced glucose-stimulated insulin secretion both

Language: Английский

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Coxsackievirus B infection invokes unique cell-type specific responses in primary human pancreatic islets

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: July 24, 2024

Coxsackievirus B (CVB) infection has long been considered an environmental factor precipitating Type 1 diabetes (T1D), autoimmune disease marked by loss of insulin-producing β cells within pancreatic islets. Previous studies have shown CVB negatively impacts islet function and viability but do not report on how virus individually affects the multiple cell types present in human primary Therefore, we hypothesized that various populations unique transcriptional responses to infection. Here, performed single-cell RNA sequencing cadaveric islets treated with either or poly(I:C), a viral mimic, for 24 48 hours. Our global analysis reveals differentially induces dynamic changes associated processes functions over time whereas poly(I:C) promotes immune response progressively increases treatment duration. At resolution, find infects all at similar rates yet cell-type specific β, α, ductal having strongest response. Sequencing functional data suggest mitochondrial respiration morphology distinct ways α cells, while also promoting generation reactive oxygen species. We observe increase expression long-noncoding

Language: Английский

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Limitations in mitochondrial programming restrain the differentiation and maturation of human stem cell-derived β cells

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: July 29, 2024

ABSTRACT Pluripotent stem cell (SC)-derived islets offer hope as a renewable source for β replacement type 1 diabetes (T1D), yet functional and metabolic immaturity may limit their long-term therapeutic potential. Here, we show that limitations in mitochondrial transcriptional programming impede the formation maturation of SC-derived (SC-β) cells. Utilizing transcriptomic profiling, assessments chromatin accessibility, phenotyping, lipidomics analyses, observed SC-β cells exhibit reduced oxidative fatty acid metabolism compared to primary human are related key networks. Surprisingly, found reductions glucose- stimulated respiration SC-islets were not associated with alterations mass, structure, or genome integrity. In contrast, limited expression targets PPARIZ PPARγ, which regulate programming, whose functions differentiation unknown. Importantly, treatment WY14643, potent agonist, induced targets, improved insulin secretion, increased both vitro following transplantation. Thus, promotes be promising target improve efforts T1D.

Language: Английский

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An INSULIN and IAPP dual reporter enables tracking of functional maturation of stem cell-derived insulin producing cells

Molecular Metabolism, Journal Year: 2024, Volume and Issue: 89, P. 102017 - 102017

Published: Aug. 24, 2024

Human embryonic stem cell (hESC; SC)-derived pancreatic β cells can be used to study diabetes pathologies and develop replacement therapies. Although current differentiation protocols yield SCβ with varying degrees of maturation, these still differ from deceased donor human in several respects. We sought a reporter line that could dynamically track functional maturation.

Language: Английский

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Depolymerizing F-actin accelerates the exit from pluripotency to enhance stem cell-derived islet differentiation

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 21, 2024

Improving generation of insulin-producing islets from human pluripotent stem cells (hPSCs) would enhance their clinical relevance for treating diabetes. Here, we demonstrate that cytoskeletal state at the onset differentiation is critical definitive endoderm formation. Depolymerizing F-actin with latrunculin A (latA) during first 24 hours facilitates rapid exit pluripotency and alters Activin/Nodal, BMP, JNK-JUN, WNT pathway signaling dynamics These changes influence downstream patterning gut tube, leading to improved pancreatic progenitor identity decreased expression markers other endodermal lineages. Continued generates containing a higher percentage β exhibit maturation, insulin secretion, ability reverse hyperglycemia. Furthermore, this latA treatment reduces enterochromaffin in final cell population corrects differentiations hPSC lines otherwise fail consistently produce islets, highlighting importance onset.

Language: Английский

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