Epigenetic biomarkers indicate islet cell death in xenotransplantation

Christopher Faulk; Kate R. Mueller; David Cheishvili; Mathia Colwell; Anne Sophie Pepin; Moshe Syzf; Bernhard J. Hering; Christopher Burlak

doi:10.1111/xen.12570

Epigenetic biomarkers indicate islet cell death in xenotransplantation

Christopher Faulk, Kate R. Mueller, David Cheishvili, Mathia Colwell, Anne Sophie Pepin, Moshe Syzf, Bernhard J. Hering, Christopher Burlak

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1 Scopus citations

Abstract

Background: Xenotransplantation of porcine islets has emerged in recent decades as a potential treatment for type 1 diabetes (T1D). Current methods of detection, indicative of successful engraftment, occur downstream of actual islet death. Epigenetic biomarkers can be detected in circulating cell-free DNA (cfDNA) to provide an earlier indication of graft dysfunction. Aims: The present study identified a biomarker of islet death using differential methylation of the insulin gene, INS, originating from β-cells in porcine islets. Materials & Methods: Pyrosequencing primers specific for porcine INS were designed to quantify hypomethylation along 12 cysteine-guanine dinucleotide (CpG) sites, including three sites in the cyclic adenosine monophosphate (cAMP) response element (CRE) binding protein 2 (CRE2) binding region of the 5′ untranslated region (UTR) and nine sites within intron 2. Results: PCR amplification of bisulfite-converted DNA combined with pyrosequencing data support the conclusion that hypomethylated porcine INS is specific to islet origin. Conclusion: Moreover, the results of this study indicate a highly specific epigenetic biomarker, capable of detecting a single islet, supporting the measurement of cfDNA as a biomarker for transplanted islet death. Defining the epigenetic characteristics of porcine-derived islets within cfDNA will be crucial to develop a better understanding of graft survival immunology for transplantation.

Original language	English (US)
Article number	e12570
Journal	Xenotransplantation
Volume	27
Issue number	2
DOIs	https://doi.org/10.1111/xen.12570
State	Published - Mar 1 2020

Bibliographical note

Funding Information:
We appreciate the contributions made by Amanda Bybee as a student in our laboratory. We gratefully acknowledge the University of Minnesota's Preclinical Research Center, directed by Dr Melanie Graham, that conducted studies in non-human primates from which retention samples were obtained. The blood, plasma, and serum samples used in this study were collected from NHP enrolled in studies funded by the Juvenile Diabetes Research Foundation (17-2013-495 and 3-SRA-2016-259-S-B) and the National Institutes of Health (1U01AI120130).

Funding Information:
We appreciate the contributions made by Amanda Bybee as a student in our laboratory. We gratefully acknowledge the University of Minnesota's Preclinical Research Center, directed by Dr Melanie Graham, that conducted studies in non‐human primates from which retention samples were obtained. The blood, plasma, and serum samples used in this study were collected from NHP enrolled in studies funded by the Juvenile Diabetes Research Foundation (17‐2013‐495 and 3‐SRA‐2016‐259‐S‐B) and the National Institutes of Health (1U01AI120130).

Publisher Copyright:
© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Keywords

DNA methylation
INS
Porcine islets
epigenetic biomarker
histone methylation
hypomethylation
insulin
xenotransplantation
β-cells

PubMed: MeSH publication types

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1111/xen.12570

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title = "Epigenetic biomarkers indicate islet cell death in xenotransplantation",

abstract = "Background: Xenotransplantation of porcine islets has emerged in recent decades as a potential treatment for type 1 diabetes (T1D). Current methods of detection, indicative of successful engraftment, occur downstream of actual islet death. Epigenetic biomarkers can be detected in circulating cell-free DNA (cfDNA) to provide an earlier indication of graft dysfunction. Aims: The present study identified a biomarker of islet death using differential methylation of the insulin gene, INS, originating from β-cells in porcine islets. Materials & Methods: Pyrosequencing primers specific for porcine INS were designed to quantify hypomethylation along 12 cysteine-guanine dinucleotide (CpG) sites, including three sites in the cyclic adenosine monophosphate (cAMP) response element (CRE) binding protein 2 (CRE2) binding region of the 5′ untranslated region (UTR) and nine sites within intron 2. Results: PCR amplification of bisulfite-converted DNA combined with pyrosequencing data support the conclusion that hypomethylated porcine INS is specific to islet origin. Conclusion: Moreover, the results of this study indicate a highly specific epigenetic biomarker, capable of detecting a single islet, supporting the measurement of cfDNA as a biomarker for transplanted islet death. Defining the epigenetic characteristics of porcine-derived islets within cfDNA will be crucial to develop a better understanding of graft survival immunology for transplantation.",

keywords = "DNA methylation, INS, Porcine islets, epigenetic biomarker, histone methylation, hypomethylation, insulin, xenotransplantation, β-cells",

author = "Christopher Faulk and Mueller, {Kate R.} and David Cheishvili and Mathia Colwell and Pepin, {Anne Sophie} and Moshe Syzf and Hering, {Bernhard J.} and Christopher Burlak",

note = "Funding Information: We appreciate the contributions made by Amanda Bybee as a student in our laboratory. We gratefully acknowledge the University of Minnesota's Preclinical Research Center, directed by Dr Melanie Graham, that conducted studies in non-human primates from which retention samples were obtained. The blood, plasma, and serum samples used in this study were collected from NHP enrolled in studies funded by the Juvenile Diabetes Research Foundation (17-2013-495 and 3-SRA-2016-259-S-B) and the National Institutes of Health (1U01AI120130). Funding Information: We appreciate the contributions made by Amanda Bybee as a student in our laboratory. We gratefully acknowledge the University of Minnesota's Preclinical Research Center, directed by Dr Melanie Graham, that conducted studies in non‐human primates from which retention samples were obtained. The blood, plasma, and serum samples used in this study were collected from NHP enrolled in studies funded by the Juvenile Diabetes Research Foundation (17‐2013‐495 and 3‐SRA‐2016‐259‐S‐B) and the National Institutes of Health (1U01AI120130). Publisher Copyright: {\textcopyright} 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.",

year = "2020",

month = mar,

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doi = "10.1111/xen.12570",

language = "English (US)",

volume = "27",

journal = "Xenotransplantation",

issn = "0908-665X",

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T1 - Epigenetic biomarkers indicate islet cell death in xenotransplantation

AU - Faulk, Christopher

AU - Mueller, Kate R.

AU - Cheishvili, David

AU - Colwell, Mathia

AU - Pepin, Anne Sophie

AU - Syzf, Moshe

AU - Hering, Bernhard J.

AU - Burlak, Christopher

N1 - Funding Information: We appreciate the contributions made by Amanda Bybee as a student in our laboratory. We gratefully acknowledge the University of Minnesota's Preclinical Research Center, directed by Dr Melanie Graham, that conducted studies in non-human primates from which retention samples were obtained. The blood, plasma, and serum samples used in this study were collected from NHP enrolled in studies funded by the Juvenile Diabetes Research Foundation (17-2013-495 and 3-SRA-2016-259-S-B) and the National Institutes of Health (1U01AI120130). Funding Information: We appreciate the contributions made by Amanda Bybee as a student in our laboratory. We gratefully acknowledge the University of Minnesota's Preclinical Research Center, directed by Dr Melanie Graham, that conducted studies in non‐human primates from which retention samples were obtained. The blood, plasma, and serum samples used in this study were collected from NHP enrolled in studies funded by the Juvenile Diabetes Research Foundation (17‐2013‐495 and 3‐SRA‐2016‐259‐S‐B) and the National Institutes of Health (1U01AI120130). Publisher Copyright: © 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

PY - 2020/3/1

Y1 - 2020/3/1

N2 - Background: Xenotransplantation of porcine islets has emerged in recent decades as a potential treatment for type 1 diabetes (T1D). Current methods of detection, indicative of successful engraftment, occur downstream of actual islet death. Epigenetic biomarkers can be detected in circulating cell-free DNA (cfDNA) to provide an earlier indication of graft dysfunction. Aims: The present study identified a biomarker of islet death using differential methylation of the insulin gene, INS, originating from β-cells in porcine islets. Materials & Methods: Pyrosequencing primers specific for porcine INS were designed to quantify hypomethylation along 12 cysteine-guanine dinucleotide (CpG) sites, including three sites in the cyclic adenosine monophosphate (cAMP) response element (CRE) binding protein 2 (CRE2) binding region of the 5′ untranslated region (UTR) and nine sites within intron 2. Results: PCR amplification of bisulfite-converted DNA combined with pyrosequencing data support the conclusion that hypomethylated porcine INS is specific to islet origin. Conclusion: Moreover, the results of this study indicate a highly specific epigenetic biomarker, capable of detecting a single islet, supporting the measurement of cfDNA as a biomarker for transplanted islet death. Defining the epigenetic characteristics of porcine-derived islets within cfDNA will be crucial to develop a better understanding of graft survival immunology for transplantation.

AB - Background: Xenotransplantation of porcine islets has emerged in recent decades as a potential treatment for type 1 diabetes (T1D). Current methods of detection, indicative of successful engraftment, occur downstream of actual islet death. Epigenetic biomarkers can be detected in circulating cell-free DNA (cfDNA) to provide an earlier indication of graft dysfunction. Aims: The present study identified a biomarker of islet death using differential methylation of the insulin gene, INS, originating from β-cells in porcine islets. Materials & Methods: Pyrosequencing primers specific for porcine INS were designed to quantify hypomethylation along 12 cysteine-guanine dinucleotide (CpG) sites, including three sites in the cyclic adenosine monophosphate (cAMP) response element (CRE) binding protein 2 (CRE2) binding region of the 5′ untranslated region (UTR) and nine sites within intron 2. Results: PCR amplification of bisulfite-converted DNA combined with pyrosequencing data support the conclusion that hypomethylated porcine INS is specific to islet origin. Conclusion: Moreover, the results of this study indicate a highly specific epigenetic biomarker, capable of detecting a single islet, supporting the measurement of cfDNA as a biomarker for transplanted islet death. Defining the epigenetic characteristics of porcine-derived islets within cfDNA will be crucial to develop a better understanding of graft survival immunology for transplantation.

KW - DNA methylation

KW - INS

KW - Porcine islets

KW - epigenetic biomarker

KW - histone methylation

KW - hypomethylation

KW - insulin

KW - xenotransplantation

KW - β-cells

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Epigenetic biomarkers indicate islet cell death in xenotransplantation

Abstract

Bibliographical note

Keywords

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UN SDGs

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