{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T10:44:28Z","timestamp":1787481868200,"version":"build-2736575974"},"reference-count":41,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2024,5,9]],"date-time":"2024-05-09T00:00:00Z","timestamp":1715212800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 AR068429"],"award-info":[{"award-number":["R01 AR068429"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["U54HD090255"],"award-info":[{"award-number":["U54HD090255"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J cachexia sarcopenia muscle"],"published-print":{"date-parts":[[2024,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Autosomal\u2010recessive mutations in <jats:italic>SPEG<\/jats:italic> (striated muscle preferentially expressed protein kinase) have been linked to centronuclear myopathy with or without dilated cardiomyopathy (CNM5). Loss of SPEG is associated with defective triad formation, abnormal excitation\u2013contraction coupling, calcium mishandling and disruption of the focal adhesion complex in skeletal muscles. To elucidate the underlying molecular pathways, we have utilized multi\u2010omics tools and analysis to obtain a comprehensive view of the complex biological processes and molecular functions.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Skeletal muscles from 2\u2010month\u2010old SPEG\u2010deficient (<jats:italic>Speg<\/jats:italic>\u2010CKO) and wild\u2010type (WT) mice were used for RNA sequencing (<jats:italic>n<\/jats:italic>\u00a0=\u00a04 per genotype) to profile transcriptomics and mass spectrometry (<jats:italic>n<\/jats:italic>\u00a0=\u00a04 for WT; <jats:italic>n<\/jats:italic>\u00a0=\u00a03 for <jats:italic>Speg<\/jats:italic>\u2010CKO mice) to profile proteomics and phosphoproteomics. In addition, interactomics was performed using the SPEG antibody on pooled muscle lysates (quadriceps, gastrocnemius and triceps) from WT and <jats:italic>Speg<\/jats:italic>\u2010CKO mice. Based on the multi\u2010omics results, we performed quantitative real\u2010time PCR, co\u2010immunoprecipitation and immunoblot to verify the findings.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We identified that SPEG interacts with myospryn complex proteins CMYA5, FSD2 and RyR1, which are critical for triad formation, and that SPEG deficiency results in myospryn complex abnormalities (protein levels decreased to 22\u00a0\u00b1\u00a03% for CMYA5 [<jats:italic>P<\/jats:italic>\u00a0&lt;\u00a00.05] and 18\u00a0\u00b1\u00a03% for FSD2 [<jats:italic>P<\/jats:italic>\u00a0&lt;\u00a00.01]). Furthermore, SPEG phosphorylates RyR1 at S2902 (phosphorylation level decreased to 55\u00a0\u00b1\u00a015% at S2902 in <jats:italic>Speg<\/jats:italic>\u2010CKO mice; <jats:italic>P<\/jats:italic>\u00a0&lt;\u00a00.05), and its loss affects JPH2 phosphorylation at multiple sites (increased phosphorylation at T161 [1.90\u00a0\u00b1\u00a00.24\u2010fold], S162 [1.61\u00a0\u00b1\u00a00.37\u2010fold] and S165 [1.66\u00a0\u00b1\u00a00.13\u2010fold]; decreased phosphorylation at S228 and S231 [39\u00a0\u00b1\u00a06%], S234 [50\u00a0\u00b1\u00a012%], S593 [48\u00a0\u00b1\u00a03%] and S613 [66\u00a0\u00b1\u00a010%]; <jats:italic>P<\/jats:italic>\u00a0&lt;\u00a00.05 for S162 and <jats:italic>P<\/jats:italic>\u00a0&lt;\u00a00.01 for other sites). On analysing the transcriptome, the most dysregulated pathways affected by SPEG deficiency included extracellular matrix\u2013receptor interaction (<jats:italic>P<\/jats:italic>\u00a0&lt;\u00a01e<jats:sup>\u221215<\/jats:sup>) and peroxisome proliferator\u2010activated receptor signalling (<jats:italic>P<\/jats:italic>\u00a0&lt;\u00a09e<jats:sup>\u221214<\/jats:sup>).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>We have elucidated the critical role of SPEG in the triad as it works closely with myospryn complex proteins (CMYA5, FSD2 and RyR1), it regulates phosphorylation levels of various residues in JPH2 and S2902 in RyR1, and its deficiency is associated with dysregulation of several pathways. The study identifies unique SPEG\u2010interacting proteins and their phosphorylation functions and emphasizes the importance of using a multi\u2010omics approach to comprehensively evaluate the molecular function of proteins involved in various genetic disorders.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jcsm.13470","type":"journal-article","created":{"date-parts":[[2024,5,10]],"date-time":"2024-05-10T06:54:09Z","timestamp":1715324049000},"page":"1003-1015","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Integrated multi\u2010omics approach reveals the role of striated muscle preferentially expressed protein kinase in skeletal muscle including its relationship with myospryn complex"],"prefix":"10.1002","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0069-8512","authenticated-orcid":false,"given":"Qifei","family":"Li","sequence":"first","affiliation":[{"name":"Division of Neonatology, Department of Pediatrics University of Miami Miller School of Medicine and Holtz Children's Hospital, Jackson Health System  Miami FL USA"},{"name":"Division of Genetics and Genomics Boston Children's Hospital, Harvard Medical School  Boston MA USA"},{"name":"The Manton Center for Orphan Disease Research Boston Children's Hospital, Harvard Medical School  Boston MA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jasmine","family":"Lin","sequence":"additional","affiliation":[{"name":"Division of Genetics and Genomics Boston Children's Hospital, Harvard Medical School  Boston MA USA"},{"name":"The Manton Center for Orphan Disease Research Boston Children's Hospital, Harvard Medical School  Boston MA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiyu","family":"Luo","sequence":"additional","affiliation":[{"name":"Division of Neonatology, Department of Pediatrics University of Miami Miller School of Medicine and Holtz Children's Hospital, Jackson Health System  Miami FL USA"},{"name":"Division of Genetics and Genomics Boston Children's Hospital, Harvard Medical School  Boston MA USA"},{"name":"The Manton Center for Orphan Disease Research Boston Children's Hospital, Harvard Medical School  Boston MA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Klaus","family":"Schmitz\u2010Abe","sequence":"additional","affiliation":[{"name":"Division of Neonatology, Department of Pediatrics University of Miami Miller School of Medicine and Holtz Children's Hospital, Jackson Health System  Miami FL USA"},{"name":"Division of Genetics and Genomics Boston Children's Hospital, Harvard Medical School  Boston MA USA"},{"name":"The Manton Center for Orphan Disease Research Boston Children's Hospital, Harvard Medical School  Boston MA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rohan","family":"Agrawal","sequence":"additional","affiliation":[{"name":"Division of Neonatology, Department of Pediatrics University of Miami Miller School of Medicine and Holtz Children's Hospital, Jackson Health System  Miami FL USA"},{"name":"Division of Genetics and Genomics Boston Children's Hospital, Harvard Medical School  Boston MA USA"},{"name":"The Manton Center for Orphan Disease Research Boston Children's Hospital, Harvard Medical School  Boston MA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa","family":"Meng","sequence":"additional","affiliation":[{"name":"Division of Genetics and Genomics Boston Children's Hospital, Harvard Medical School  Boston MA USA"},{"name":"The Manton Center for Orphan Disease Research Boston Children's Hospital, Harvard Medical School  Boston MA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Behzad","family":"Moghadaszadeh","sequence":"additional","affiliation":[{"name":"Division of Genetics and Genomics Boston Children's Hospital, Harvard Medical School  Boston MA USA"},{"name":"The Manton Center for Orphan Disease Research 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