Acute Ischemia Induced by High Density Culture Increases Cytokine Expression and Diminishes the Function and Viability of Highly Purified Human Islets of Langerhans

Kate E. Smith, Amy C. Kelly, Catherine G. Min, Craig S. Weber, Fiona M. McCarthy, Leah V. Steyn, Vasudeo Badarinarayana, Brett B. Stanton, Jennifer P. Kitzmann, Peter Strop, Angelika C. Gruessner, Ronald M. Lynch, Sean W. Limesand, Klearchos K. Papas

Research output: Research - peer-reviewArticle

  • 2 Citations

Abstract

BACKGROUND: Encapsulation devices have the potential to enable cell based insulin replacement therapies (such as human islet or stem cell derived β cell transplantation) without immunosuppression. However, reasonably sized encapsulation devices promote ischemia due to high β cell densities creating prohibitively large diffusional distances for nutrients. It is hypothesized that even acute ischemic exposure will compromise the therapeutic potential of cell based insulin replacement. In this study, the acute effects of high-density ischemia were investigated in human islets to develop a detailed profile of early ischemia induced changes and targets for intervention. METHODS: Human islets were exposed in a pairwise model simulating high density encapsulation to normoxic or ischemic culture for 12 hours, after which viability and function were measured. RNA sequencing (RNAseq) was conducted to assess transcriptome-wide changes in gene expression. RESULTS: Islet viability after acute ischemic exposure was reduced compared to normoxic culture conditions (p<0.01). Insulin secretion was also diminished, with ischemic β cells losing their insulin secretory response to stimulatory glucose levels (p<0.01). RNAseq revealed 657 differentially expressed genes following ischemia, with many that are associated with increased inflammatory and hypoxia-response signaling and decreased nutrient transport and metabolism. CONCLUSIONS: In order for cell based insulin replacement to be applied as a treatment for type 1 diabetes, oxygen and nutrient delivery to β cells will need to be maintained. We demonstrate that even brief ischemic exposure such as would be experienced in encapsulation devices damages islet viability and β cell function, and leads to increased inflammatory signaling.This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.

LanguageEnglish (US)
JournalTransplantation
DOIs
StateAccepted/In press - Mar 3 2017

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Islets of Langerhans
Ischemia
Insulin
Cytokines
Food
Equipment and Supplies
Therapeutics
RNA Sequence Analysis
Cell Transplantation
Licensure
Type 1 Diabetes Mellitus
Transcriptome
Immunosuppression
Cell Survival
Stem Cells
Cell Count
Oxygen
Gene Expression
Glucose
Genes

ASJC Scopus subject areas

  • Transplantation

Cite this

Acute Ischemia Induced by High Density Culture Increases Cytokine Expression and Diminishes the Function and Viability of Highly Purified Human Islets of Langerhans. / Smith, Kate E.; Kelly, Amy C.; Min, Catherine G.; Weber, Craig S.; McCarthy, Fiona M.; Steyn, Leah V.; Badarinarayana, Vasudeo; Stanton, Brett B.; Kitzmann, Jennifer P.; Strop, Peter; Gruessner, Angelika C.; Lynch, Ronald M.; Limesand, Sean W.; Papas, Klearchos K.

In: Transplantation, 03.03.2017.

Research output: Research - peer-reviewArticle

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abstract = "BACKGROUND: Encapsulation devices have the potential to enable cell based insulin replacement therapies (such as human islet or stem cell derived β cell transplantation) without immunosuppression. However, reasonably sized encapsulation devices promote ischemia due to high β cell densities creating prohibitively large diffusional distances for nutrients. It is hypothesized that even acute ischemic exposure will compromise the therapeutic potential of cell based insulin replacement. In this study, the acute effects of high-density ischemia were investigated in human islets to develop a detailed profile of early ischemia induced changes and targets for intervention. METHODS: Human islets were exposed in a pairwise model simulating high density encapsulation to normoxic or ischemic culture for 12 hours, after which viability and function were measured. RNA sequencing (RNAseq) was conducted to assess transcriptome-wide changes in gene expression. RESULTS: Islet viability after acute ischemic exposure was reduced compared to normoxic culture conditions (p<0.01). Insulin secretion was also diminished, with ischemic β cells losing their insulin secretory response to stimulatory glucose levels (p<0.01). RNAseq revealed 657 differentially expressed genes following ischemia, with many that are associated with increased inflammatory and hypoxia-response signaling and decreased nutrient transport and metabolism. CONCLUSIONS: In order for cell based insulin replacement to be applied as a treatment for type 1 diabetes, oxygen and nutrient delivery to β cells will need to be maintained. We demonstrate that even brief ischemic exposure such as would be experienced in encapsulation devices damages islet viability and β cell function, and leads to increased inflammatory signaling.This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.",
author = "Smith, {Kate E.} and Kelly, {Amy C.} and Min, {Catherine G.} and Weber, {Craig S.} and McCarthy, {Fiona M.} and Steyn, {Leah V.} and Vasudeo Badarinarayana and Stanton, {Brett B.} and Kitzmann, {Jennifer P.} and Peter Strop and Gruessner, {Angelika C.} and Lynch, {Ronald M.} and Limesand, {Sean W.} and Papas, {Klearchos K.}",
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AU - Smith,Kate E.

AU - Kelly,Amy C.

AU - Min,Catherine G.

AU - Weber,Craig S.

AU - McCarthy,Fiona M.

AU - Steyn,Leah V.

AU - Badarinarayana,Vasudeo

AU - Stanton,Brett B.

AU - Kitzmann,Jennifer P.

AU - Strop,Peter

AU - Gruessner,Angelika C.

AU - Lynch,Ronald M.

AU - Limesand,Sean W.

AU - Papas,Klearchos K.

PY - 2017/3/3

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N2 - BACKGROUND: Encapsulation devices have the potential to enable cell based insulin replacement therapies (such as human islet or stem cell derived β cell transplantation) without immunosuppression. However, reasonably sized encapsulation devices promote ischemia due to high β cell densities creating prohibitively large diffusional distances for nutrients. It is hypothesized that even acute ischemic exposure will compromise the therapeutic potential of cell based insulin replacement. In this study, the acute effects of high-density ischemia were investigated in human islets to develop a detailed profile of early ischemia induced changes and targets for intervention. METHODS: Human islets were exposed in a pairwise model simulating high density encapsulation to normoxic or ischemic culture for 12 hours, after which viability and function were measured. RNA sequencing (RNAseq) was conducted to assess transcriptome-wide changes in gene expression. RESULTS: Islet viability after acute ischemic exposure was reduced compared to normoxic culture conditions (p<0.01). Insulin secretion was also diminished, with ischemic β cells losing their insulin secretory response to stimulatory glucose levels (p<0.01). RNAseq revealed 657 differentially expressed genes following ischemia, with many that are associated with increased inflammatory and hypoxia-response signaling and decreased nutrient transport and metabolism. CONCLUSIONS: In order for cell based insulin replacement to be applied as a treatment for type 1 diabetes, oxygen and nutrient delivery to β cells will need to be maintained. We demonstrate that even brief ischemic exposure such as would be experienced in encapsulation devices damages islet viability and β cell function, and leads to increased inflammatory signaling.This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.

AB - BACKGROUND: Encapsulation devices have the potential to enable cell based insulin replacement therapies (such as human islet or stem cell derived β cell transplantation) without immunosuppression. However, reasonably sized encapsulation devices promote ischemia due to high β cell densities creating prohibitively large diffusional distances for nutrients. It is hypothesized that even acute ischemic exposure will compromise the therapeutic potential of cell based insulin replacement. In this study, the acute effects of high-density ischemia were investigated in human islets to develop a detailed profile of early ischemia induced changes and targets for intervention. METHODS: Human islets were exposed in a pairwise model simulating high density encapsulation to normoxic or ischemic culture for 12 hours, after which viability and function were measured. RNA sequencing (RNAseq) was conducted to assess transcriptome-wide changes in gene expression. RESULTS: Islet viability after acute ischemic exposure was reduced compared to normoxic culture conditions (p<0.01). Insulin secretion was also diminished, with ischemic β cells losing their insulin secretory response to stimulatory glucose levels (p<0.01). RNAseq revealed 657 differentially expressed genes following ischemia, with many that are associated with increased inflammatory and hypoxia-response signaling and decreased nutrient transport and metabolism. CONCLUSIONS: In order for cell based insulin replacement to be applied as a treatment for type 1 diabetes, oxygen and nutrient delivery to β cells will need to be maintained. We demonstrate that even brief ischemic exposure such as would be experienced in encapsulation devices damages islet viability and β cell function, and leads to increased inflammatory signaling.This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-No Derivatives License 4.0 (CCBY-NC-ND), where it is permissible to download and share the work provided it is properly cited. The work cannot be changed in any way or used commercially without permission from the journal.

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