{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T16:01:39Z","timestamp":1785427299517,"version":"3.56.0"},"reference-count":35,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T00:00:00Z","timestamp":1784073600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Masone Family"},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"crossref","award":["HA 556161976"],"award-info":[{"award-number":["HA 556161976"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"crossref"}]},{"award":["HA 556161976"],"award-info":[{"award-number":["HA 556161976"]}],"id":[{"id":"https:\/\/ror.org\/018mejw64","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background: ACTA2 encodes smooth muscle alpha-actin and is one of the most common genetic causes of inherited non-syndromic thoracic aortic aneurysm and dissection. Although murine models have provided important mechanistic insight, complementary vertebrate systems may enable more rapid in vivo phenotyping and future therapeutic screening. Methods: Utilizing a CRISPR\/Cas9-based approach, we developed zebrafish acta2 mutant models and assessed phenotypes at larval and adult stages. At 3 days post-fertilization, larvae were screened by brightfield microscopy for edema, axis defects, hemorrhage, and a composite endpoint (any phenotype) under basal conditions and after exposure to 0.2 mM epinephrine. Phenotype occurrence was summarized as group-specific proportions across a 2 \u00d7 2 design defined by genotype (wild-type vs. acta2-deficient mutant) and treatment (DMSO control vs. epinephrine), with prespecified pairwise comparisons using Pearson\u2019s chi-squared tests. Pooled fish-level logistic regression models were also fit for each endpoint. Adult follow-up was performed by genotyping all available acta2 fish at the facility and tracking staggered tank-level cohorts longitudinally. Results: Initial gross morphologic assessment did not reveal overt external phenotypic differences between heterozygous or homozygous acta2 mutant larvae and wild-type controls. In larval analyses, the mutant genotype was associated with a greater burden of adverse phenotypes than treatment exposure. For the composite endpoint, mutant larvae demonstrated higher proportions than normal larvae under both control (28.3% vs. 17.4%, p = 0.007) and epinephrine conditions (26.9% vs. 17.4%, p = 0.023), whereas epinephrine did not significantly alter composite phenotype frequency within either genotype. Similar genotype-associated trends were observed for axis defects and hemorrhage. In pooled logistic regression, acta2 deficiency was associated with increased odds of axis defects (OR 2.64, 95% CI 1.48\u20134.89, p = 0.001), hemorrhage (OR 2.40, 95% CI 1.09\u20135.68, p = 0.036), and the composite endpoint (OR 1.88, 95% CI 1.19\u20133.00, p = 0.008), whereas epinephrine exposure did not demonstrate a consistent independent effect across endpoints. Adult staggered follow-up showed greater attrition in homozygous mutant cohorts than in wild-type or heterozygous cohorts, with aggregated losses of 24%, 12%, and 3%, respectively. Conclusions: This preliminary zebrafish acta2 model demonstrated that acta2 deficiency is associated with increased adverse larval phenotype burden and suggested reduced long-term persistence of homozygous mutant fish in adulthood. The strongest signal in the current study was genotype-associated phenotype burden rather than a robust epinephrine-dependent effect. These findings support the feasibility of zebrafish-based ACTA2 phenotyping while highlighting the need for more specific vascular endpoints, refined longitudinal follow-up, and future variant-specific modeling.<\/jats:p>","DOI":"10.3390\/genes17070808","type":"journal-article","created":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T08:08:34Z","timestamp":1784189314000},"page":"808","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Preliminary Zebrafish Model of ACTA2 Deficiency Reveals Increased Larval Phenotype Burden and Suggests Reduced Adult Mutant Survival"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1666-199X","authenticated-orcid":false,"given":"Mohammad A.","family":"Zafar","sequence":"first","affiliation":[{"name":"Aortic Institute at Yale-New Haven, Yale University School of Medicine, New Haven, CT 06519, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-9991-7981","authenticated-orcid":false,"given":"Lisa C.","family":"Harling","sequence":"additional","affiliation":[{"name":"Aortic Institute at Yale-New Haven, Yale University School of Medicine, New Haven, CT 06519, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2063-1067","authenticated-orcid":false,"given":"Yupeng","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Political Science & Economics, Rowan University, Glassboro, NJ 08028, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-6272-925X","authenticated-orcid":false,"given":"Nafiye B.","family":"Celik","sequence":"additional","affiliation":[{"name":"Aortic Institute at Yale-New Haven, Yale University School of Medicine, New Haven, CT 06519, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sandip K.","family":"Mukherjee","sequence":"additional","affiliation":[{"name":"Aortic Institute at Yale-New Haven, Yale University School of Medicine, New Haven, CT 06519, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0366-8503","authenticated-orcid":false,"given":"John","family":"Rizzo","sequence":"additional","affiliation":[{"name":"Program in Public Health, Stony Brook University, Stony Brook, NY 11794, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"Prendergast","sequence":"additional","affiliation":[{"name":"Yale Zebrafish Research Core, Yale University School of Medicine, New Haven, CT 06511, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6255-8139","authenticated-orcid":false,"given":"John A.","family":"Elefteriades","sequence":"additional","affiliation":[{"name":"Aortic Institute at Yale-New Haven, Yale University School of Medicine, New Haven, CT 06519, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4579","DOI":"10.1093\/eurheartj\/ehad148","article-title":"Fate of the unoperated ascending thoracic aortic aneurysm: Three-decade experience from the Aortic Institute at Yale University","volume":"44","author":"Wu","year":"2023","journal-title":"Eur. 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