- •We differentiated intra-islet substructures by their proteome profiles in situ.
- •We demonstrated significant functional differences between vasculature types.
- •Novel differences in innervation and colocalization of nerves were observed.
- •We demonstrated a depth of coverage useful for predictive binding discovery assays.
- •We identified pathways with these techniques that could aid drug characterizations.
Abbreviations:ABC (ammonium bicarbonate), FAIMS (high-field asymmetric waveform ion mobility spectrometry), LCM (laser capture microdissection), nanoPOTS (nanodroplet processing in one-pot for trace samples)
Experimental Design and Statistical Rationale
Collection of Pancreata From Rats and Imaging of Pancreatic Islet Vasculature
Laser Capture Microdissection
NanoPOTS-Based Protein Extraction and Digestion
Depth of Proteome Coverage
Distinct Global Proteome Profiles
Feature-Specific Protein Markers
Enriched Signatures Characteristic of Innervation and Neuron-Vasculature-Islet Crosstalk Within the Islet Inner Vasculature
- Alvarsson A.
- Jimenez-Gonzalez M.
- Li R.
- Rosselot C.
- Tzavaras N.
- Wu Z.
- et al.
- Alvarsson A.
- Jimenez-Gonzalez M.
- Li R.
- Rosselot C.
- Tzavaras N.
- Wu Z.
- et al.
Ligandable Cysteine-–Containing Proteins Toward Drug–Target Interactome at the Sub-Islet Scale
Limitations of the Current Study
- Jongmin Woo S.M.W.
- Aguilera-Vazquez V.
- Sontag R.L.
- Moore R.J.
- Markillie L.M.
- Mehta H.S.
- et al.
Conflict of interest
Funding and additional information
- Supplemental Figures
- Supplementary Tables Protein and Peptide Quant rev3
- Single PSM protein spectra
- Islet beta cell mass in diabetes and how it relates to function, birth, and death.Ann. N. Y Acad. Sci. 2013; 1281: 92-105
- Molecular and functional profiling of human islets: from heterogeneity to human phenotypes.Diabetologia. 2020; 63: 2095-2101
- Single-cell transcriptome profiling of human pancreatic islets in health and type 2 diabetes.Cell Metab. 2016; 24: 593-607
- Heterogeneity of the human pancreatic islet.Diabetes. 2019; 68: 1230-1239
- Islet microenvironment, modulated by vascular endothelial growth factor-A signaling, promotes beta cell regeneration.Cell Metab. 2014; 19: 498-511
- Vascular endothelium in diabetes.Am. J. Physiol. Ren. Physiol. 2017; 312: F266-F275
- Improved in vivo imaging method for individual islets across the mouse pancreas reveals a heterogeneous insulin secretion response to glucose.Sci. Rep. 2021; 11: 603
- Pancreatic islet plasticity: Interspecies comparison of islet architecture and composition.Islets. 2010; 2: 135-145
- Islet architecture: a comparative study.Islets. 2009; 1: 129-136
- Human islets contain four distinct subtypes of beta cells.Nat. Commun. 2016; 711756
- Vascular heterogeneity between native rat pancreatic islets is responsible for differences in survival and revascularisation post transplantation.Diabetologia. 2015; 58: 132-139
- Impact of islet architecture on beta-cell heterogeneity, plasticity and function.Nat. Rev. Endocrinol. 2016; 12: 695-709
- Single-cell multi-omics analysis of human pancreatic islets reveals novel cellular states in type 1 diabetes.Nat. Metab. 2022; 4: 284-299
- Single-cell transcriptomics of the human endocrine pancreas.Diabetes. 2016; 65: 3028-3038
- Interrogating islets in health and disease with single-cell technologies.Mol. Metab. 2017; 6: 991-1001
- A single-cell transcriptome atlas of the human pancreas.Cell Syst. 2016; 3: 385-394.e383
- Single-cell transcriptomes identify human islet cell signatures and reveal cell-type-specific expression changes in type 2 diabetes.Genome Res. 2017; 27: 208-222
- RNA sequencing of single human islet cells reveals type 2 diabetes genes.Cell Metab. 2016; 24: 608-615
- Single cell peptide heterogeneity of rat islets of Langerhans.ACS Chem. Biol. 2016; 11: 2588-2595
- Proteomic profiling of human islets collected from frozen pancreata using laser capture microdissection.J. Proteomics. 2017; 150: 149-159
- Characterization of the mouse pancreatic islet proteome and comparative analysis with other mouse tissues.J. Proteome Res. 2008; 7: 3114-3126
- Discovery of novel glucose-regulated proteins in isolated human pancreatic islets using LC-MS/MS-based proteomics.J. Proteome Res. 2012; 11: 3520-3532
- Nanodroplet processing platform for deep and quantitative proteome profiling of 10-100 mammalian cells.Nat. Commun. 2018; 9: 882
- Quantitative proteomic analysis of single pancreatic islets.Proc. Natl. Acad. Sci. U. S. A. 2009; 106: 18902-18907
- Mechanistic investigations of test article-induced pancreatic toxicity at the endocrine-exocrine interface in the rat.Toxicol. Pathol. 2014; 42: 229-242
- Spatially resolved proteome mapping of laser capture microdissected tissue with automated sample transfer to nanodroplets.Mol. Cell Proteomics. 2018; 17: 1864-1874
- Automated mass spectrometry imaging of over 2000 proteins from tissue sections at 100-mum spatial resolution.Nat. Commun. 2020; 11: 8
- Three-dimensional feature matching improves coverage for single-cell proteomics based on ion mobility filtering.Cell Syst. 2022; 13: 426-434.e424
- Automated coupling of nanodroplet sample preparation with liquid chromatography-mass spectrometry for high-throughput single-cell proteomics.Anal. Chem. 2020; 92: 10588-10596
- Ultrasensitive single-cell proteomics workflow identifies >1000 protein groups per mammalian cell.Chem. Sci. 2021; 12: 1001-1006
- Robust, sensitive, and quantitative single-cell proteomics based on ion mobility filtering.bioRxiv. 2021; ([preprint])https://doi.org/10.1101/2021.01.30.428333
- The MaxQuant computational platform for mass spectrometry-based shotgun proteomics.Nat. Protoc. 2016; 11: 2301-2319
- DAnTE: a statistical tool for quantitative analysis of -omics data.Bioinformatics. 2008; 24: 1556-1558
- DAVID bioinformatics resources: expanded annotation database and novel algorithms to better extract biology from large gene lists.Nucl. Acids Res. 2007; 35: W169-W175
- GOrilla: a tool for discovery and visualization of enriched GO terms in ranked gene lists.BMC Bioinform. 2009; 10: 48
- A human protein atlas for normal and cancer tissues based on antibody proteomics.Mol. Cell Proteomics. 2005; 4: 1920-1932
- Proteomics. Tissue-based map of the human proteome.Science. 2015; 3471260419
- The human pancreatic islet transcriptome: expression of candidate genes for type 1 diabetes and the impact of pro-inflammatory cytokines.PLoS Genet. 2012; 8e1002552
- Annexins family: insights into their functions and potential role in pathogenesis of sarcoidosis.J. Transl. Med. 2016; 14: 89
- Distribution and ultrastructure of the autonomic nerves in the mouse pancreas.Microsc. Res. Tech. 1997; 37: 399-406
- Macro- and micro-domains in the endocrine pancreas.Diabetes. 1982; 31: 538-565
- A 3D atlas of the dynamic and regional variation of pancreatic innervation in diabetes.Sci. Adv. 2020; 6https://doi.org/10.1126/sciadv.aaz9124
- Remodelling sympathetic innervation in rat pancreatic islets ontogeny.BMC Dev. Biol. 2009; 9: 34
- Beta cells are important for islet innervation: evidence from purified rat islet-cell grafts.Diabetologia. 1996; 39: 54-59
- Noninvasive in vivo model demonstrating the effects of autonomic innervation on pancreatic islet function.Proc. Natl. Acad. Sci. U. S. A. 2012; 109: 21456-21461
- Innervation patterns of autonomic axons in the human endocrine pancreas.Cell Metab. 2011; 14: 45-54
- The human protein atlas: a spatial map of the human proteome.Protein Sci. 2018; 27: 233-244
- The synaptic proteins neurexins and neuroligins are widely expressed in the vascular system and contribute to its functions.Proc. Natl. Acad. Sci. U. S. A. 2009; 106: 20782-20787
- Neuronal and vascular interactions.Annu. Rev. Neurosci. 2015; 38: 25-46
- Cross-talk between blood vessels and neural progenitors in the developing brain.Neuronal. Signal. 2018; 2NS20170139
- Signalling between microvascular endothelium and cardiomyocytes through neuregulin.Cardiovasc. Res. 2014; 102: 194-204
- Neural control of the endocrine pancreas.Best Pract. Res. Clin. Endocrinol. Metab. 2014; 28: 745-756
- Single-cell transcriptomics from human pancreatic islets: sample preparation matters.Biol. Met. Protoc. 2020; 5: bpz019
- Single-cell transcriptomes reveal characteristic features of human pancreatic islet cell types.EMBO Rep. 2016; 17: 178-187
- An improved boosting to amplify signal with isobaric labeling (iBASIL) strategy for precise quantitative single-cell proteomics.Mol. Cell Proteomics. 2020; 19: 828-838
- High-throughput and high-efficiency sample preparation for single-cell proteomics using a nested nanowell chip.bioRxiv. 2021; ([preprint])https://doi.org/10.1101/2021.02.17.431689
- High-throughput and high-efficiency sample preparation for single-cell proteomics using a nested nanowell chip.Nat. Commun. 2021; 12: 6246
- PDV: an integrative proteomics data viewer.Bioinformatics. 2019; 35: 1249-1251
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