Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets

Blodgett, David M.; Nowosielska, Anetta; Afik, Shaked; Pechhold, Susanne; Cura, Anthony J.; Kennedy, Norman J.; Kim, Soyoung; Küçükural, Alper; Davis, Roger J.; Kent, Sally C.; Greiner, Dale L.; Garber, Manuel; Harlan, David M.; Diiorio, Philip J.

doi:10.2337/db15-0039

Cited by 273 publications

(410 citation statements)

References 41 publications

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“…High-throughput analysis of mRNA abundance with RNA sequencing (RNA-seq) has confirmed MT1 and MT2 expression in human islets, though equal abundance of transcript for each was reported in alpha and beta cells [107]. In the pancreatic islet, melatonin regulates a bevy of cellular activity and gene expression [108][109][110][111].…”

Section: The Pancreatic Response To Melatoninmentioning

confidence: 99%

Chronomedicine and type 2 diabetes: shining some light on melatonin

et al. 2016

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In mammals, the circadian timing system drives rhythms of physiology and behaviour, including the daily rhythms of feeding and activity. The timing system coordinates temporal variation in the biochemical landscape with changes in nutrient intake in order to optimise energy balance and maintain metabolic homeostasis. Circadian disruption (e.g. as a result of shift work or jet lag) can disturb this continuity and increase the risk of cardiometabolic disease. Obesity and metabolic disease can also disturb the timing and amplitude of the clock in multiple organ systems, further exacerbating disease progression. As our understanding of the synergy between the timing system and metabolism has grown, an interest has emerged in the development of novel clock-targeting pharmaceuticals or nutraceuticals for the treatment of metabolic dysfunction. Recently, the pineal hormone melatonin has received some attention as a potential chronotherapeutic drug for metabolic disease. Melatonin is well known for its sleep-promoting effects and putative activity as a chronobiotic drug, stimulating coordination of biochemical oscillations through targeting the internal timing system. Melatonin affects the insulin secretory activity of the pancreatic beta cell, hepatic glucose metabolism and insulin sensitivity. Individuals with type 2 diabetes mellitus have lower night-time serum melatonin levels and increased risk of comorbid sleep disturbances compared with healthy individuals. Further, reduced melatonin levels, and mutations and/or genetic polymorphisms of the melatonin receptors are associated with an increased risk of developing type 2 diabetes. Herein we review our understanding of molecular clock control of glucose homeostasis, detail the influence of circadian disruption on glucose metabolism in critical peripheral tissues, explore the contribution of melatonin signalling to the aetiology of type 2 diabetes, and discuss the pros and cons of melatonin chronopharmacotherapy in disease management.

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Section: The Pancreatic Response To Melatoninmentioning

confidence: 99%

Chronomedicine and type 2 diabetes: shining some light on melatonin

et al. 2016

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“…ZnT6 mRNA was detected in the islets of Langerhans [34], its functional heterodimerization with ZnT5 has been described [35], and its presence in purified β cells from both mouse [36] and human [37,38] confirmed by massive parallel sequencing (RNASeq).…”

Section: Znt Familymentioning

confidence: 94%

“…ZnT3 (SLC30A3) protein has been reported in the islets, and in the pancreatic β cell line INS-1E but not in a corresponding pancreatic α-cell line, α-TC6 [40][41][42]. While ZnT3 mRNA was reported in purified human β cells in one study [38], levels were zero, or at the limit of detection, in samples from several subjects in another [37]. ZnT3 mRNA was similarly barely 5 detected by RNASeq in purified mouse β cells [36], in line with our findings in mouse islets [34].…”

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confidence: 95%

Zinc and diabetes

Chabosseau

Rutter

2016

Archives of Biochemistry and Biophysics

139

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Zn 2+ ions are essential for the normal processing and storage of insulin and altered pancreatic insulin content is associated with all forms of diabetes mellitus. Work of the past decade has identified variants in the human SLC30A8 gene, encoding the zinc transporter ZnT8 which is expressed highly selectively on the secretory granule of pancreatic islet β and α cells, as affecting the risk of Type 2 Diabetes. Here, we review the regulation and roles of Zn 2+ ions in islet cells, the mechanisms through which SLC30A8 variants might affect glucose homeostasis and diabetes risk, and the novel technologies including recombinant targeted zinc probes and knockout mice which have been developed to explore these questions. Abbreviation ISG: Insulin Secreting Granules; T2D: Type 2 Diabetes; T1D: Type 1 Diabetes; 2 Diabetes mellitus is a common metabolic disease, affecting approximately 9% of the adult population worldwide. It is characterized by high circulating glucose levels over a prolonged period of time which leads to long-term health complications [1]. Type 1 Diabetes (T1D) involves the autoimmune destruction of insulin-secreting β cells while Type 2 Diabetes (T2D) is linked to both a decrease of insulin release and deficient hormone action on targeted organs [2].The links among Zn 2+ , diabetes and insulin were established in the 1930s, a decade after the discovery of the hormone, when a study showed crystallized insulin contains Zn 2+ and that Zn 2+ , along with other metal ions, could reversibly trigger insulin crystallization [3]. Zinc was later demonstrated to prolong insulin action when co-injected with the hormone [4].These early studies, as well as much more recent findings showing association between T2D risk and the inheritance of gene variants encoding a critical β cell Zn 2+ transporter, ZnT8[5] have generated considerable interest in the role of Zn 2+ in diabetes aetiology and as a possible therapeutic target [6]. Zinc and the diabetic patientScott and Fisher were the first to report a direct link between zinc and diabetes in patients.While assessing the insulin content in the pancreas of diabetic patients compared to nondiabetic cadavers, they showed that in the former group zinc content was reduced by 75 % [7].Epidemiological studies also demonstrated that diabetes and whole body zinc status are associated [8][9][10][11]. In T1D and T2D patients, serum zinc levels are significantly decreased [9][10][11], this being associated with an increased zinc urinary loss [8].Zinc may have important anti-oxidant properties, as it acts as a cofactor of the superoxide dismutase enzyme which regulates the detoxification of reactive oxygen species, regulating the expression and protecting against the oxidative stress induced by chronic hyperglycaemia (for review, [12]). Furthermore, zinc inhibits alpha-ketoglutarate-dependent mitochondrial respiration suggesting that Zn 2+ can interfere with mitochondrial antioxidant production and may also stimulate production of reactive oxygen [13].Zinc supplementation h...

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“…This system is ideal for two reasons. First, all previous descriptions of islet composition have relied on known markers of cells (Pechhold et al 2009;Dorrell et al 2011;Hrvatin et al 2014;Blodgett et al 2015) or sequenced a relatively small number of cells (70) . inDrop allows the profiling of thousands of cells, and hence, rare cell populations of even <1% are likely to be well represented.…”

Section: Single-cell Islet Sequencing Reveals a Complex Cellular Compmentioning

confidence: 99%

“…We used nondiabetic BBDRs as islet donors and isolated and dissociated the islets into single-cell suspensions as previously described (Blodgett et al 2015). Cells were flowed through the inDrop system to obtain single-cell RNAseq libraries, which were then pooled together for sequencing as previously described ( Fig.…”

Section: Single-cell Islet Sequencing Reveals a Complex Cellular Compmentioning

confidence: 99%

End Sequence Analysis Toolkit (ESAT) expands the extractable information from single-cell RNA-seq data

et al. 2016

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RNA-seq protocols that focus on transcript termini are well suited for applications in which template quantity is limiting. Here we show that, when applied to end-sequencing data, analytical methods designed for global RNA-seq produce computational artifacts. To remedy this, we created the End Sequence Analysis Toolkit (ESAT). As a test, we first compared endsequencing and bulk RNA-seq using RNA from dendritic cells stimulated with lipopolysaccharide (LPS). As predicted by the telescripting model for transcriptional bursts, ESAT detected an LPS-stimulated shift to shorter 3 ′ -isoforms that was not evident by conventional computational methods. Then, droplet-based microfluidics was used to generate 1000 cDNA libraries, each from an individual pancreatic islet cell. ESAT identified nine distinct cell types, three distinct β-cell types, and a complex interplay between hormone secretion and vascularization. ESAT, then, offers a much-needed and generally applicable computational pipeline for either bulk or single-cell RNA end-sequencing.

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Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets

Cited by 273 publications

References 41 publications

Chronomedicine and type 2 diabetes: shining some light on melatonin

Chronomedicine and type 2 diabetes: shining some light on melatonin

Zinc and diabetes

End Sequence Analysis Toolkit (ESAT) expands the extractable information from single-cell RNA-seq data

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