Novel Observations From Next-Generation RNA Sequencing of Highly Purified Human Adult and Fetal Islet Cell Subsets
Open accessAbstract
Understanding distinct gene expression patterns of normal adult and developing fetal human pancreatic α- and β-cells is crucial for developing stem cell therapies, islet regeneration strategies, and therapies designed to increase β-cell function in patients with diabetes (type 1 or 2). Toward that end, we have developed methods to highly purify α-, β-, and δ-cells from human fetal and adult pancreata by intracellular staining for the cell-specific hormone content, sorting the subpopulations by flow cytometry, and, using next-generation RNA sequencing, we report the detailed transcriptomes of fetal and adult α- and β-cells. We observed that human islet composition was not influenced by age, sex, or BMI, and transcripts for inflammatory gene products were noted in fetal β-cells. In addition, within highly purified adult glucagon-expressing α-cells, we observed surprisingly high insulin mRNA expression, but not insulin protein expression. This transcriptome analysis from highly purified islet α- and β-cell subsets from fetal and adult pancreata offers clear implications for strategies that seek to increase insulin expression in type 1 and type 2 diabetes.
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IAPP (amylin) is expressed at levels 1% and 10% of that of insulin in human and mouse β-cells, respectively.
“IAPP (amylin) is expressed at levels 1% and 10% of that of insulin in human (68, 475) and mouse β-cells, respectively (Appendix Figure 1A).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section, “Established and Enriched Transcripts in Highly Purified α- and β-Cell Populations” (page unavailable), identifies IAPP and INS as robustly expressed human β-cell genes. It does not give a 1% IAPP-to-insulin expression ratio. The cited paper also does not establish the mouse β-cell ratio; in the citing sentence, that portion is attached to an appendix figure rather than this citation. (LLM, conf=0.78)
Experimental — accuracy not yet measuredThe Na-K ATPase of both mouse and human β-cells is predominantly composed of α1-, β1- and β3-subunits.
“A tetrameric complex of two α- and two β-subunits, the Na-K ATPase of both mouse and human β-cells is predominantly composed of α1-, β1- and β3-subunits (56, 68).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched α- and β-cell transcripts lists several β-cell genes but does not discuss Na-K ATPase subunits. Searches of our copy for Na-K ATPase, ATPase, ATP1A1, ATP1B1, and ATP1B3 found no passage establishing that α1, β1, and β3 predominate in human or mouse β-cells. (LLM, conf=0.75)
Experimental — accuracy not yet measuredIn contrast to mouse β-cells, the delayed rectifying K+ current in human β-cells is carried principally by Kv2.2 (KCNB2) and Kv1.6 (KCNA6) channels.
“In contrast to mouse β-cells, the delayed rectifying K+ current in human β-cells is carried principally by Kv2.2 (KCNB2) and Kv1.6 (KCNA6) channels (Appendix Figure 5A), which are expressed at high levels in the human β-cell and encode inactivating TEA-sensitive voltage-gated K+ channels (68, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched transcripts reports β-cell expression of KCNQ2, but does not address Kv2.2 (KCNB2) or Kv1.6 (KCNA6). Searches for both genes and for delayed-rectifier currents and TEA sensitivity found no passage establishing which channels principally carry the human β-cell current or comparing it with mouse β-cells. Page numbers were not available in our copy. (LLM, conf=0.76)
Experimental — accuracy not yet measuredHuman β-cells mainly express SCN8A (Nav1.6), accounting for 40% of Na+ channel transcripts, with the remainder being SCN9A (Nav1.7), SCN3A (Nav1.3) and SCN2A (Nav1.2).
“Whereas mouse β-cells mainly express Scn9a (Nav1.7), SCN8A (Nav1.6) accounts for 40% of Na+ channel transcripts in human β-cells (68, 475) with the remainder being SCN9A (Nav1.7), SCN3A (Nav1.3) and SCN2A (Nav1.2) (see Appendix Figure 4).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched α- and β-cell transcripts reports β-cell gene expression but does not give the claimed sodium-channel breakdown. Searches for SCN8A, Nav1.6, and the 40% figure found no passage in our copy stating the proportions or identifying SCN9A, SCN3A, and SCN2A as the remainder; page numbers were not available. (LLM, conf=0.72)
Experimental — accuracy not yet measuredLike their mouse counterparts, human β-cells predominantly express SK3 (KCNN3), with other SKs being expressed at 10-fold lower levels.
“Like their mouse counterparts, human β-cells predominantly express SK3 (KCNN3) with other SKs being expressed at 10-fold lower levels (68, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched α- and β-cell transcripts names KCNQ2 and KCND3, but does not report KCNN3 or compare expression of SK channels. Searches of our copy for KCNN3, SK3, and the claimed 10-fold difference found no passage addressing that comparison. (LLM, conf=0.72)
Experimental — accuracy not yet measured
In human β-cells, ITPR3 is expressed at particularly high levels, approximately 10-fold greater than ITPR1 or ITPR2.
“In human β-cells, ITPR3 is expressed at particularly high levels (≈10-fold greater than ITPR1 or ITPR2) (68, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched β-cell transcripts lists several genes but does not mention ITPR3, ITPR1, or ITPR2. Searches for those genes and for inositol trisphosphate receptor comparisons found no passage in our copy supporting the approximately 10-fold expression difference. (LLM, conf=0.72)
Experimental — accuracy not yet measuredBoth mouse and human β-cells express Hcn/HCN2-4.
“Both mouse (3, 146) and human (68, 475) β-cells express Hcn/HCN2-4.”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched β-cell transcripts lists several human β-cell genes but does not mention HCN2, HCN3, or HCN4; the Discussion also does not address their expression. Searches for those gene names and hyperpolarization-activated channels found no relevant passage in our copy, so it cannot verify the human β-cell portion attributed to this source. (LLM, conf=0.72)
Experimental — accuracy not yet measuredExpression of GABAA receptor β-subunits in human β-cells is much higher than α-subunits and is entirely accounted for by GABRB3.
“Expression of the β-subunits is much higher (Appendix Figure 18B) and is entirely accounted for by GABRB3 (68, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched transcripts in purified α- and β-cell populations lists several β-cell genes, but does not report GABAA receptor subunit expression. Searches for GABRB3, other GABAA subunit genes, and a β-versus-α subunit comparison found no passage in our copy addressing the claim; no page numbers were available. (LLM, conf=0.76)
Experimental — accuracy not yet measuredGABA transporters GAT1-3 are only expressed at low levels in mouse and human β-cells.
“However, these are only expressed at low levels in mouse and human β-cells alike (3, 68, 146, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on established and enriched transcripts in purified α- and β-cells lists several β-cell genes, but does not give expression levels for GAT1–3. Searches of our copy for GABA transporters, GAT1–3, and their gene names found no passage addressing their expression in human or mouse β-cells. (LLM, conf=0.72)
Experimental — accuracy not yet measuredSLC17A9 expression in human β-cells is almost undetectable.
“Although SLC17A9 expression in human β-cells is almost undetectable (68, 475), stimulation of insulin secretion is still associated with ATP release (347).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section discusses gene-expression profiles in purified human β-cells, but the passages in our copy do not report SLC17A9 expression. Searches for “SLC17A9” and for low expression of a vesicular nucleotide transporter in β-cells found no passage that establishes the claim; no page numbers were provided. (LLM, conf=0.72)
Experimental — accuracy not yet measuredBoth human and mouse β-cells express Slc6a17/SLC6a17, which encodes a synaptic vesicle glycine transporter.
“Both human and mouse β-cells also express Slc6a17/SLC6a17 (3, 68, 146, 475) that encodes a synaptic vesicle glycine transporter (502).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched β-cell transcripts lists several genes but does not mention SLC6A17. Searches of our copy for SLC6A17 and glycine transporter found no passage establishing its expression in human or mouse β-cells. The transporter description is attached to a different citation in the citing sentence. (LLM, conf=0.72)
Experimental — accuracy not yet measuredGlycine is cleared from interstitium by plasmalemmal glycine transporter GlyT1 (SLC6A9), which cotransports Na+.
“Following its release, glycine is cleared from the interstitium by the plasmalemmal glycine transporter GlyT1 (SLC6A9) (68, 475), which cotransports Na+.”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The cited paper’s Methods and Results passages concern RNA sequencing and protein expression in purified human islet cells, not glycine transport. Searches for GlyT1, SLC6A9, glycine clearance, and sodium cotransport found no passage addressing the claim. (LLM, conf=0.76)
Experimental — accuracy not yet measuredVglut2 (SLC17A6) is expressed in human β-cells.
“Similarly, Vglut2 (SLC17A6) is expressed in human β-cells ((68) and glutamate may therefore be co-released with insulin from β-cells (205).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched transcripts in purified human β-cells lists several expressed genes but does not mention SLC17A6 or VGLUT2. Searches for both names and for vesicular glutamate transporter found no passage in our copy establishing its expression in human β-cells. (LLM, conf=0.72)
Experimental — accuracy not yet measuredExpression of functional GABAA receptors is very low in human α-cells.
“However, it should be noted that the expression of functional GABAA receptors is very low in human α-cells (89), an observation confirmed at the mRNA level (68, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The citation to this paper is attached to the claimed mRNA-level confirmation. The Results section on enriched α- and β-cell transcripts discusses genes such as GCG and ARX, but the passages found by searching for GABA, GABAA receptors, and receptor subunits do not report GABAA-receptor mRNA levels in human α-cells. (LLM, conf=0.70)
Experimental — accuracy not yet measuredMouse and human β-cells express CAPS1 and low amounts of CAPS2.
“Mouse and human β-cells express CAPS1 (Cadps/CADPS) and low amounts of CAPS2 (Cadps2/CADPS2) (3, 68, 146, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched β-cell transcripts lists several genes but does not mention CADPS or CADPS2. Searches for CADPS/CADPS2, CAPS1/CAPS2, and their descriptive names found no passage in our copy establishing their expression levels in human or mouse β-cells. The Results refer to supplementary tables, but the cited passages do not provide those values. (LLM, conf=0.73)
Experimental — accuracy not yet measuredIn human β-cells, only SYN1 is expressed, whereas mouse β-cells express both Syn1 and Syn2.
“In human β-cells, only SYN1 is expressed, whereas mouse β-cells express both Syn1 and Syn2 (3, 68, 146, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on enriched β-cell transcripts lists several human β-cell genes but does not address SYN1 or SYN2. Searches for SYN1, SYN2, and the claimed human–mouse comparison found no passage in our copy establishing their expression in either species. (LLM, conf=0.74)
Experimental — accuracy not yet measuredDoc2A and Doc2B are expressed at low levels in mouse and human β-cells.
“These are both expressed at low levels in mouse and human β-cells (3, 68, 146, 475).”
From Pancreatic β-Cell Electrical Activity and Insulin Secretion: Of Mice and Men (2018) · cites this paper as [68]
The Results section on established and enriched transcripts in purified human α- and β-cells reports expression of several genes, but does not mention Doc2A or Doc2B. Searches for both gene names and their low-level expression in mouse and human β-cells found no passage addressing the claim in our copy. (LLM, conf=0.72)
Experimental — accuracy not yet measured
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- Authors
- Blodgett, D. M., Nowosielska, A., Afik, S., Pechhold, S., Cura, A. J., Kennedy, N. J., Kim, S., Kucukural, A., Davis, R. J., Kent, S. C., Greiner, D. L., Garber, M. G., Harlan, D. M., & diIorio, P.
- Affiliations
- University of Massachusetts Chan Medical School (US)
- Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, Worcester, MA
- Howard Hughes Medical Institute (US)
- Corresponding author
- Harlan, David M.
- ORCID
- Venue
- Diabetes
- Publisher
- American Diabetes Association
- Published
- 2015-04-30
- Volume
- 64
- Issue
- 9
- Pages
- 3172-3181
- ISSN
- 0012-1797 (ISSN-L)
- 1939-327X
- PMC
- PMC4542439
- OpenAlex
- W2103920106
- Cited by
- 330
- References
- 42
- Citations by year
- 2026: 11 · 2025: 21 · 2024: 22 · 2023: 40 · 2022: 30 · 2021: 42 · 2020: 32 · 2019: 30 · 2018: 36 · 2017: 36 · 2016: 23 · 2015: 5
- FWCI
- 33.56
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Diabetes (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- The Journal of the American Medical Association (JAMA) Network (American Medical Association) (submittedVersion)
- Funders
- National Institute of Diabetes and Digestive and Kidney Diseases (U01 DK089572, UC4 DK104218)
- Topics
- Health Sciences › Medicine › Surgery › Pancreatic function and diabetes
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Diabetes and associated disorders
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Single-cell and spatial transcriptomics
- Keywords
- pancreatic islet cells, alpha cells, pancreatic beta cells, delta cells, fetal pancreas, adult pancreas, cell purification, transcriptome, glucagon, insulin mRNA expression, inflammatory gene expression, islet regeneration, stem cell therapy, type 1 diabetes, type 2 diabetes
- MeSH terms
- Adolescent, Adult, Child, Preschool, Female, Fetus, Humans, Islets of Langerhans, Male, Middle Aged, Pregnancy, Pregnancy Trimester, Second, RNA, Sequence Analysis, RNA, Gene Expression Regulation, Developmental, Somatostatin-Secreting Cells, Gene Expression Profiling, Glucagon-Secreting Cells, Insulin-Secreting Cells, Young Adult
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