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We developed an engineered human micro-heart tissue (\u03bcHT) combining iPSC-derived cardiomyocytes (CM) with defined primary cardiac fibroblast (cFB) fractions and mechanically modulated afterload. This design allowed us to test how stromal context and pro-fibrotic signaling shape HCM pathogenesis in micro-engineered heart tissues made from iPSCs harboring an HCM-linked frameshift variant in MYBPC3 (MYBPC3fs). Under low afterload, genotype-associated differences in total active force were limited at 0% and 5% added cFB but became apparent at 25% added cFB. However, high afterload markedly amplified MYBPC3fs \u03bcHT hypercontractility, with tissues generating greater total active force at both 5% and 25% added cFB. We also observed afterload- and cFB-dependent remodeling of Ca2+ handling, indicating that excitation\u2013contraction coupling in MYBPC3fs tissues is modulated by combinatorial actions of mechanical stress and the stromal environment. Despite heightened contractile output, the MYBPC3fs \u03bcHT exhibited blunted improvements in Z-disc alignment with increasing cFB content, whereas cardiomyocytes within the isogenic control \u03bcHT showed progressive structural ordering as cFB density increased. Finally, the MYBPC3fs \u03bcHT exhibited a greater response to TGF-\u03b2, leading to exaggerated increases in tissue-level resting tension and \u03b1-smooth muscle actin expression. Our findings provide important insights into the pathophysiologic pathways leading to adverse outcomes in clinical HCM and a foundation for future studies of cardiomyocyte\u2013fibroblast interactions in HCM.<\/jats:p>","DOI":"10.1063\/5.0336866","type":"journal-article","created":{"date-parts":[[2026,9,14]],"date-time":"2026-09-14T13:52:30Z","timestamp":1789393950000},"update-policy":"https:\/\/doi.org\/10.1063\/aip-crossmark-policy-page","source":"Crossref","is-referenced-by-count":2,"title":["Biomechanical stress unmasks a fibroblast-dependent hypercontractile-disarray phenotype in MYBPC3 truncation HCM"],"prefix":"10.1063","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0781-896X","authenticated-orcid":false,"given":"Huanzhu","family":"Jiang","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering, Washington University School of Engineering 1 , St. Louis, Missouri 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