{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"medRxiv"}],"indexed":{"date-parts":[[2026,1,16]],"date-time":"2026-01-16T12:58:18Z","timestamp":1768568298928,"version":"3.49.0"},"posted":{"date-parts":[[2024,3,19]]},"group-title":"Allergy and Immunology","reference-count":9,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2024,3,19]]},"abstract":"Abstract<\/jats:title>\n \n Background<\/jats:title>\n \n Phospholipase C\u03b32 (PLC\u03b32) is an important signaling molecule that receives and transmits signals from various cell surface receptors in most hematopoietic lineages. Variants of\n PLCG2<\/jats:italic>\n cause PLC\u03b32-associated immune dysregulation (PLAID), a family of conditions that are classified by mutational effect. PLAID with cold urticaria (CU-PLAID) is caused by in-frame deletions of\n PLCG2<\/jats:italic>\n that are dominant negative at physiologic temperatures but become spontaneously active at sub-physiologic temperatures.\n <\/jats:p>\n <\/jats:sec>\n \n Objective<\/jats:title>\n \n To identify genetic lesions that cause PLAID by combining RNA sequencing of full-length\n PLCG2<\/jats:italic>\n with whole genome sequencing.\n <\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n We studied nine probands with antibody deficiency and a positive evaporative cooling test, together with two known CU-PLAID patients and three healthy subjects. Illumina sequencing was performed on full-length\n PLCG2<\/jats:italic>\n cDNA synthesized from peripheral blood mononuclear cell RNA and whole genome sequencing was used to identify genetic lesions. Novel alternate transcripts were overexpressed in the\n Plcg2-<\/jats:italic>\n deficient DT40 cell overexpression system. ERK phosphorylation was quantified by flow cytometry with and without BCR crosslinking.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Two probands expressed novel alternative transcripts of\n PLCG2<\/jats:italic>\n with in-frame deletions. The first, expressing\n PLCG2<\/jats:italic>\n without exons 18-19, carried a splice site mutation in intron 19. The second, expressing\n PLCG2<\/jats:italic>\n without exons 19-22, carried a 14kb\n de novo<\/jats:italic>\n deletion of\n PLCG2<\/jats:italic>\n . DT40 cells overexpressing the exon 18-19 or exon 19-22 deletions failed to phosphorylate ERK in response to BCR crosslinking.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n \n In addition to autosomal dominant genomic deletions,\n de novo<\/jats:italic>\n deletions and splice site mutations of\n PLCG2<\/jats:italic>\n can also cause CU-PLAID. All of these can be identified by cDNA-based sequencing.\n <\/jats:p>\n <\/jats:sec>\n \n Capsule Summary<\/jats:title>\n \n By identifying both the first\n de novo<\/jats:italic>\n and splice site variants to cause\n PLCG2<\/jats:italic>\n -associated immune dysregulation with cold urticaria (CU-PLAID), we demonstrate the diagnostic utility of\n PLCG2<\/jats:italic>\n -specific RNA-sequencing.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.1101\/2024.03.16.24304180","type":"posted-content","created":{"date-parts":[[2024,3,19]],"date-time":"2024-03-19T15:25:33Z","timestamp":1710861933000},"source":"Crossref","is-referenced-by-count":0,"title":["Splice site and\n de novo<\/i>\n mutations can cause mixed dominant negative\/gain of function\n PLCG2<\/i>\n -associated immune dysregulation with cold urticaria (CU-PLAID)"],"prefix":"10.64898","author":[{"given":"Sophia R.","family":"Chou","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexis C.","family":"Bailey","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kathleen","family":"Baysac","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew J.","family":"Oler","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joshua D.","family":"Milner","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3322-4089","authenticated-orcid":false,"given":"Michael J.","family":"Ombrello","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2024032110101714000_2024.03.16.24304180v1.1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbc.2021.100905"},{"issue":"1","key":"2024032110101714000_2024.03.16.24304180v1.2","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1016\/j.jaci.2023.08.036","article-title":"PLCG2-associated immune dysregulation (PLAID) comprises broad and distinct clinical presentations related to functional classes of genetic variants","volume":"153","year":"2024","journal-title":"J Allergy Clin Immunol"},{"key":"2024032110101714000_2024.03.16.24304180v1.3","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1102140"},{"key":"2024032110101714000_2024.03.16.24304180v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/j.ajhg.2012.08.006"},{"issue":"1","key":"2024032110101714000_2024.03.16.24304180v1.5","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1016\/j.jaci.2023.09.002","article-title":"Human PLCG2 haploinsufficiency results in a novel natural killer cell immunodeficiency","volume":"153","year":"2024","journal-title":"J Allergy Clin Immunol"},{"issue":"6","key":"2024032110101714000_2024.03.16.24304180v1.6","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1001\/jamadermatol.2014.5641","article-title":"Distinct Cutaneous Manifestations and Cold-Induced Leukocyte Activation Associated With PLCG2 Mutations","volume":"151","year":"2015","journal-title":"JAMA Dermatol"},{"key":"2024032110101714000_2024.03.16.24304180v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaci.2009.09.035"},{"key":"2024032110101714000_2024.03.16.24304180v1.8","doi-asserted-by":"publisher","DOI":"10.1038\/s41588-018-0071-6"},{"key":"2024032110101714000_2024.03.16.24304180v1.9","doi-asserted-by":"crossref","unstructured":"Lee Y , Wessel AW , Xu J , et al. Genetically programmed alternative splicing of NEMO mediates an autoinflammatory disease phenotype. J Clin Invest. 2022 Mar 15;132(6).","DOI":"10.1172\/JCI128808"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2024.03.16.24304180","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T19:55:43Z","timestamp":1768506943000},"score":1,"resource":{"primary":{"URL":"http:\/\/medrxiv.org\/lookup\/doi\/10.1101\/2024.03.16.24304180"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,19]]},"references-count":9,"URL":"https:\/\/doi.org\/10.1101\/2024.03.16.24304180","relation":{},"subject":[],"published":{"date-parts":[[2024,3,19]]},"subtype":"preprint"}}