Proteomic profiling of human islets collected from frozen pancreata using laser capture microdissection

Zhang, Lina; Lanzoni, Giacomo; Battarra, Matteo; Inverardi, Luca; Zhang, Qibin

doi:10.1016/j.jprot.2016.09.002

Cited by 19 publications

(14 citation statements)

References 55 publications

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“…Two previous studies have utilized LCM and mass spectrometry based proteomics to analyze differential protein expression in human islets obtained under different conditions and pathological states [ 56 – 57 ]. Unlike our current study, Nishida et al [ 56 ], performed qualitative proteomic analysis to evaluate protein expression using the islets isolated from formalin-fixed paraffin-embedded pancreatic tissues from three autopsied cases of enterovirus-associated fulminant type 1 diabetes (FT1DM) with extensive insulitis compared to five autopsied non-diabetic men.…”

Section: Resultsmentioning

confidence: 99%

“…Unlike our current study, Nishida et al [ 56 ], performed qualitative proteomic analysis to evaluate protein expression using the islets isolated from formalin-fixed paraffin-embedded pancreatic tissues from three autopsied cases of enterovirus-associated fulminant type 1 diabetes (FT1DM) with extensive insulitis compared to five autopsied non-diabetic men. On the other hand, Zhang et al [ 57 ], recently performed comparative quantitative proteomic analyses of enzyme isolated cultured islets, and laser capture microdissected human islets and acinar tissue from fresh-frozen pancreas sections from three non-diabetic cadaveric donors. Unlike these two previous reports, our current proteomics data is obtained from 3 groups of disease stratified phenotype matched comprehensively annotated pancreas tissue samples from nPOD.…”

Section: Resultsmentioning

confidence: 99%

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Comparative quantitative proteomic analysis of disease stratified laser captured microdissected human islets identifies proteins and pathways potentially related to type 1 diabetes

Nyalwidhe¹,

Grzesik²,

Burch³

et al. 2017

PLoS ONE

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Type 1 diabetes (T1D) is a chronic inflammatory disease that is characterized by autoimmune destruction of insulin-producing pancreatic beta cells. The goal of this study was to identify novel protein signatures that distinguish Islets from patients with T1D, patients who are autoantibody positive without symptoms of diabetes, and from individuals with no evidence of disease. High resolution high mass accuracy label free quantitative mass spectrometry analysis was applied to islets isolated by laser capture microdissection from disease stratified human pancreata from the Network for Pancreatic Organ Donors with Diabetes (nPOD), these included donors without diabetes, donors with T1D-associated autoantibodies in the absence of diabetes, and donors with T1D. Thirty-nine proteins were found to be differentially regulated in autoantibody positive cases compared to the no-disease group, with 25 upregulated and 14 downregulated proteins. For the T1D cases, 63 proteins were differentially expressed, with 24 upregulated and 39 downregulated, compared to the no disease controls. We have identified functional annotated enriched gene families and multiple protein-protein interaction clusters of proteins are involved in biological and molecular processes that may have a role in T1D. The proteins that are upregulated in T1D cases include S100A9, S100A8, REG1B, REG3A and C9 amongst others. These proteins have important biological functions, such as inflammation, metabolic regulation, and autoimmunity, all of which are pathways linked to the pathogenesis of T1D. The identified proteins may be involved in T1D development and pathogenesis. Our findings of novel proteins uniquely upregulated in T1D pancreas provides impetus for further investigations focusing on their expression profiles in beta cells/ islets to evaluate their role in the disease pathogenesis. Some of these molecules may be novel therapeutic targets T1D.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Comparative quantitative proteomic analysis of disease stratified laser captured microdissected human islets identifies proteins and pathways potentially related to type 1 diabetes

Nyalwidhe¹,

Grzesik²,

Burch³

et al. 2017

PLoS ONE

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“…There are no studies showing ALK3 expression at the protein level, be it by western blot, proteomics, or immunostaining. In fact, ALK3 has not been detected in proteomic analysis of either human ( Zhang et al, 2017 ) or murine ( Waanders et al, 2009 ) isolated islets. By sorting ALK3 bright+ cells (the only fraction that shows responsiveness to BMP-7), we selectively exclude any residual β cells that may have been carried over in the hNEPT.…”

Section: Discussionmentioning

confidence: 99%

P2RY1/ALK3-Expressing Cells within the Adult Human Exocrine Pancreas Are BMP-7 Expandable and Exhibit Progenitor-like Characteristics

et al. 2018

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SUMMARYTreatment of human pancreatic non-endocrine tissue with Bone Morphogenetic Protein 7 (BMP-7) leads to the formation of glucose-responsive β-like cells. Here, we show that BMP-7 acts on extrainsular cells expressing PDX1 and the BMP receptor activin-like kinase 3 (ALK3/BMPR1A). In vitro lineage tracing indicates that ALK3+ cell populations are multipotent. PDX1+/ALK3+ cells are absent from islets but prominently represented in the major pancreatic ducts and pancreatic duct glands. We identified the puri-nergic receptor P2Y1 (P2RY1) as a surrogate surface marker for PDX1. Sorted P2RY1+/ALK3bright+ cells form BMP-7-expandable colonies characterized by NKX6.1 and PDX1 expression. Unlike the negative fraction controls, these colonies can be differentiated into multiple pancreatic lineages upon BMP-7 withdrawal. RNA-seq further corroborates the progenitor-like nature of P2RY1+/ALK3bright+ cells and their multilineage differentiation potential. Our studies confirm the existence of progenitor cells in the adult human pancreas and suggest a specific anatomical location within the ductal and glandular networks.

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“…This method has the advantage of using frozen tissue, with protein able to be extracted from only the target cell under microscope observation. 34 Previous reports showed that expression of lysosomal enzymes (MDH1, PC and ACLY) involved in glucose sensing tend to be different in human and rat b cells. 35 A proteome analysis focusing on glucose-responsive insulin production, which is a cellular physiological function specific to human pancreatic b cells, was also reported.…”

Section: Discussionmentioning

confidence: 99%

A proteome analysis of pig pancreatic islets and exocrine tissue by liquid chromatography with tandem mass spectrometry

Nakashima

Miyagi-Shiohira

Kobayashi

et al. 2017

Islets

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Liquid chromatography with tandem mass spectrometry (LC-MS/MS) is a proteome analysis method, and the shotgun analysis by LC-MS/MS comprehensively identifies proteins from tissues and cells with high resolving power. In this study, we analyzed the protein expression in pancreatic tissue by LC-MS/MS. Islets isolated from porcine pancreata (purity ≥95%) and exocrine tissue (purity ≥99%) were used in this study. LC-MS/MS showed that 13 proteins were expressed in pancreatic islets only (Group I), 43 proteins were expressed in both islets and exocrine tissue (Group I&E), and 102 proteins were expressed in exocrine tissue only (Group E). Proteins involved in islet differentiation and cell proliferation were identified in Group I (e.g. CLUS, CMGA, MIF). In addition, various functional proteins (e.g. SCG2, TBA1A) were identified in islet by using the new method of 'principal component analysis (PCA)'. However, the function of such proteins on islets remains unclear. EPCAM was identified in Group E. Group E was found to include proteins involved in clinical inflammatory diseases such as pancreatitis (e.g. CBPA1, CGL, CYTB, ISK1 and PA21B). Many of these identified proteins were reported less frequently in previous studies, and HS71B, NEC2, PRAF3 and SCG1 were newly detected in Group I while CPNS1, DPEP1, GANAB, GDIB, GGT1, HSPB1, ICTL, VILI, MUTA, NDKB, PTGR1, UCHL3, VAPB and VINC were newly detected in Group E. These results show that comprehensive expression analysis of proteins by LC-MS/MS is useful as a method to investigate new factors constructing cellular component, biological process, and molecular function.

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Proteomic profiling of human islets collected from frozen pancreata using laser capture microdissection

Cited by 19 publications

References 55 publications

Comparative quantitative proteomic analysis of disease stratified laser captured microdissected human islets identifies proteins and pathways potentially related to type 1 diabetes

Comparative quantitative proteomic analysis of disease stratified laser captured microdissected human islets identifies proteins and pathways potentially related to type 1 diabetes

P2RY1/ALK3-Expressing Cells within the Adult Human Exocrine Pancreas Are BMP-7 Expandable and Exhibit Progenitor-like Characteristics

A proteome analysis of pig pancreatic islets and exocrine tissue by liquid chromatography with tandem mass spectrometry

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