{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T20:05:11Z","timestamp":1785441911438,"version":"3.56.0"},"posted":{"date-parts":[[2026,7,28]]},"group-title":"Physiology","reference-count":62,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T00:00:00Z","timestamp":1785196800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,7,28]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Cardiometabolic heart failure with preserved ejection fraction (HFpEF) is associated with systemic and cardiac inflammation and diastolic dysfunction. A-kinase anchoring protein 6\u03b2 (AKAP6\u03b2) is a scaffold protein located at the cardiomyocyte outer nuclear membrane that promotes pathological cardiac remodeling via the recruitment of multiple regulatory proteins including protein kinases. In mice, adeno-associated virus (AAV) mediated expression of a peptide based upon a kinase binding domain (KBD) within AKAP6\u03b2 inhibited the development of heart failure due to chronic pressure overload. Whether KBD expression can also inhibit the development of cardiometabolic heart failure is unknown, and if so, the mechanism of KBD action in HFpEF has yet to be explored.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>The efficacy of a cardiotropic self-complementary AAV gene therapy that expresses the AKAP6\u03b2 KBD peptide (AAV9sc.KBD) was tested in a female Ossabaw swine model of cardiometabolic syndrome and HFpEF. Single nucleus and bulk RNA sequencing of swine heart tissue and immunoprecipitation-mass spectrometry, live cell imaging, and biochemical assays using primary rat cardiomyocytes were employed to study KBD mechanism of action.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    AAV9sc.KBD inhibited the development of diastolic dysfunction and heart failure in the Ossabaw model, without negatively impacting systolic function. The improvement in cardiac phenotype was associated with decreased T-cell myocardial infiltrates and partial reversal of pathological gene expression. An unbiased interactome study revealed that the KBD peptide binds Ca\n                    <jats:sup>2+<\/jats:sup>\n                    \/calmodulin-dependent protein kinase II (CaMKII), identifying CaMKII as a new AKAP6\u03b2 binding partner. Perinuclear CaMKII activity detected by live cell imaging required AKAP6\u03b2 expression and was inhibited by KBD expression. In addition, the CaMKII substrate Inhibitor of NF-\u03baB Kinase \u03b2 (IKK\u03b2) bound AKAP6\u03b2. IKK phosphorylation in the Ossabaw model and in myocytes was inhibited by KBD expression, and NF-\u03baB nuclear translocation in myocytes was dependent upon AKAP6\u03b2-CaMKII protein complex formation. AAV9sc.KBD treatment inhibited cardiomyocyte NF-\u03baB-dependent gene expression in the Ossabaw model.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>Regulated by perinuclear AKAP6\u03b2-CaMKII signalosomes, NF-\u03baB pro-inflammatory gene expression in cardiomyocytes participates in a positive feedback loop with cardiac inflammation promoting HFpEF. Proof-of-concept is provided in a large animal model that gene therapy-based cardiomyocyte expression of the KBD peptide will prevent cardiac dysfunction in cardiometabolic syndrome.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Clinical Perspective<\/jats:title>\n                  <jats:sec>\n                    <jats:title>What is new<\/jats:title>\n                    <jats:list list-type=\"bullet\">\n                      <jats:list-item>\n                        <jats:p>The cardiomyocyte-selective gene therapy AAV9sc.KBD, which targets signalosomes organized by the scaffold protein AKAP6\u03b2, is shown to inhibit myocardial T-cell infiltration and improve cardiac structure and function in a large animal model of cardiometabolic HFpEF.<\/jats:p>\n                      <\/jats:list-item>\n                      <jats:list-item>\n                        <jats:p>The AKAP6\u03b2 KBD peptide is shown to bind and inhibit the function of CaMKII.<\/jats:p>\n                      <\/jats:list-item>\n                      <jats:list-item>\n                        <jats:p>CaMKII and IKK\u03b2 are shown to participate in perinuclear AKAP6\u03b2 signalosomes, where they regulate activation of the NF-\u03baB pro-inflammatory gene regulatory pathway.<\/jats:p>\n                      <\/jats:list-item>\n                    <\/jats:list>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Clinical implications:<\/jats:title>\n                    <jats:list list-type=\"bullet\">\n                      <jats:list-item>\n                        <jats:p>Proof-of-concept for a novel strategy for the treatment of HFpEF is provided, intracellular expression by a cardiomyocyte-selective gene therapy vector of an inhibitory peptide, which will inhibit compartmentalized intracellular signal transduction.<\/jats:p>\n                      <\/jats:list-item>\n                      <jats:list-item>\n                        <jats:p>In conjunction with previous studies in small rodents, the new data obtained in Ossabaw swine support clinical translation of the AAV9sc.KBD gene therapy.<\/jats:p>\n                      <\/jats:list-item>\n                    <\/jats:list>\n                  <\/jats:sec>\n                <\/jats:sec>","DOI":"10.64898\/2026.07.23.740435","type":"posted-content","created":{"date-parts":[[2026,7,28]],"date-time":"2026-07-28T16:50:22Z","timestamp":1785257422000},"source":"Crossref","is-referenced-by-count":0,"title":["Gene therapy targeting of AKAP6\u03b2-CaMKII signalosomes improves myocardial inflammation and heart failure in a swine model of cardiometabolic syndrome"],"prefix":"10.64898","author":[{"given":"Darla L.","family":"Tharp","sequence":"first","affiliation":[{"name":"Department of Pathobiology and Integrative Biomedical Sciences, University of Missouri-Columbia, Columbia, MO 65211"},{"name":"NextGen Precision Health, 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