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Epigenetic biomarkers indicate islet cell death in xenotransplantation
Author(s) -
Faulk Christopher,
Mueller Kate R.,
Cheishvili David,
Colwell Mathia,
Pepin AnneSophie,
Syzf Moshe,
Hering Bernhard J.,
Burlak Christopher
Publication year - 2020
Publication title -
xenotransplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.052
H-Index - 61
eISSN - 1399-3089
pISSN - 0908-665X
DOI - 10.1111/xen.12570
Subject(s) - xenotransplantation , epigenetics , biology , islet , dna methylation , biomarker , transplantation , cpg site , microbiology and biotechnology , cancer research , gene , genetics , medicine , gene expression , insulin , endocrinology
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.