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Affected genes include PRKACA, GNAS, PRKAR1A, and CTNNB1.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>This work aims to expand our understanding of the prevalence of somatic mutations in CPAs from patients with overt Cushing syndrome (OCS) and \u201csubclinical\u201d mild autonomous cortisol excess (MACE), with an immunohistochemistry (IHC)\u2012guided targeted amplicon sequencing approach using formalin-fixed paraffin-embedded (FFPE) tissue.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We analyzed FFPE adrenal tissue from 77 patients (n\u2005=\u200512 men, 65 women) with either OCS (n\u2005=\u200532) or MACE (n\u2005=\u200545). Using IHC for 17\u03b1-hydroxylase\/17,20-lyase (CYP17A1) and 3\u03b2-hydroxysteroid dehydrogenase (HSD3B2), we identified 78 CPAs (32 OCS CPAs and 46 MACE CPAs). Genomic DNA was isolated from the FFPE CPAs and subjected to targeted amplicon sequencing for identification of somatic mutations.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Somatic mutations were identified in 71.8% (56\/78) of the CPAs. While PRKACA was the most frequently mutated gene in OCS CPAs (14\/32, 43.8%), somatic genetic aberrations in CTNNB1 occurred in 56.5% (26\/46) of the MACE CPAs. Most GNAS mutations were observed in MACE CPAs (5\/7, 71.4%). No mutations were observed in PRKAR1A. In addition to the known mutations, we identified one previously unreported mutation in PRKACA. Two patients with MACE harbored 2 adjacent tumors within the same adrenal gland - one patient had 2 CPAs, and the other patient had a CPA and an aldosterone-producing adenoma (identified by IHC for aldosterone synthase).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>A comprehensive FFPE IHC-guided gene-targeted sequencing approach identified somatic mutations in 71.8% of the CPAs. OCS CPAs demonstrated a distinct mutation profile compared to MACE CPAs.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1210\/clinem\/dgab682","type":"journal-article","created":{"date-parts":[[2021,9,17]],"date-time":"2021-09-17T14:40:34Z","timestamp":1631889634000},"page":"e594-e603","source":"Crossref","is-referenced-by-count":41,"title":["Targeted Mutational Analysis of Cortisol-Producing Adenomas"],"prefix":"10.1210","volume":"107","author":[{"given":"Juilee","family":"Rege","sequence":"first","affiliation":[{"name":"Department of Molecular and Integrative Physiology, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessie","family":"Hoxie","sequence":"additional","affiliation":[{"name":"Department of Molecular and Integrative Physiology, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chia-Jen","family":"Liu","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Morgan N","family":"Cash","sequence":"additional","affiliation":[{"name":"University of Kentucky College of Medicine , Lexington, Kentucky ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James M","family":"Luther","sequence":"additional","affiliation":[{"name":"Division of Clinical Pharmacology, Vanderbilt University Medical Center , Nashville, Tennessee 37232 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lan","family":"Gellert","sequence":"additional","affiliation":[{"name":"Department of Pathology, Microbiology and Immunology, Vanderbilt University Medical Center , Nashville, Tennessee 37232 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adina F","family":"Turcu","sequence":"additional","affiliation":[{"name":"Division of Metabolism, Endocrinology, and Diabetes, Department of Internal Medicine, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2262-0011","authenticated-orcid":false,"given":"Tobias","family":"Else","sequence":"additional","affiliation":[{"name":"Division of Metabolism, Endocrinology, and Diabetes, Department of Internal Medicine, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas J","family":"Giordano","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]},{"name":"Rogel Cancer Center, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]},{"name":"Michigan Center for Translational Pathology, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aaron M","family":"Udager","sequence":"additional","affiliation":[{"name":"Department of Pathology, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]},{"name":"Rogel Cancer Center, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]},{"name":"Michigan Center for Translational Pathology, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5557-8406","authenticated-orcid":false,"given":"William E","family":"Rainey","sequence":"additional","affiliation":[{"name":"Department of Molecular and Integrative Physiology, University of Michigan , Ann Arbor, Michigan 48109 ,","place":["USA"]},{"name":"Division of Metabolism, Endocrinology, 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