479 Effects of extracellular matrix on pacemaking cardiomyocyte function
Objectives/Goals: The extracellular matrix (ECM) of the sinoatrial node (SAN) is critical for maintaining automaticity in hiPSC-derived pacemaking cardiomyocytes (PCMs) under cyclic strain. We aim to determine the ECM ligands responsible for cell-ECM mediated mechanotransduction and the resulting ph...
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| Format: | Article |
| Language: | English |
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Cambridge University Press
2025-04-01
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| Series: | Journal of Clinical and Translational Science |
| Online Access: | https://www.cambridge.org/core/product/identifier/S2059866124010707/type/journal_article |
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| author | Brian Howard Regan Smithers Kaitlin Van Brusselen Hillary K.J. Kao Deborah K. Lieu |
| author_facet | Brian Howard Regan Smithers Kaitlin Van Brusselen Hillary K.J. Kao Deborah K. Lieu |
| author_sort | Brian Howard |
| collection | DOAJ |
| description | Objectives/Goals: The extracellular matrix (ECM) of the sinoatrial node (SAN) is critical for maintaining automaticity in hiPSC-derived pacemaking cardiomyocytes (PCMs) under cyclic strain. We aim to determine the ECM ligands responsible for cell-ECM mediated mechanotransduction and the resulting phenotype in PCMs. Methods/Study Population: HiPSCs are differentiated to PCM and replated on substrate with 5 or 15 kPa PDMS that are coated with 5 or 25 ug/cm of either collagen I or fibronectin at sub-confluent density to restrict junction engagement to only costameres. Then, PCM are subjected to 10% cyclic mechanical strain at 1 Hz for 48 hours, with static culture as control. PCMs from all conditions are subsequently fixed and stained for cardiomyocyte-specific troponin T (TnT), pacemaking HCN4 channel, and pro-pacemaking transcription factors (Shox2, Isl1, Tbx3, Tbx18). Additionally, PCM cell size will also be assessed. Results/Anticipated Results: Considering the amount of hypertrophy and myofilament in CMs correlates with mechanical strain, we expect a reduced degree of mechanotransduction in hiPSC-PCM on collagen I with a stiffness 15 kPa to induce smaller cell size with fewer myofilament and an upregulation of HCN4 and pro-pacemaking transcription factors than those on 5 kPa and those on fibronectin of either 5 or 15 kPa after cyclic strain. This is because COL1 is reported to have a lower signaling threshold but a limited sensitivity to force which contributes to the diminished mechanotransduction signaling. Discussion/Significance of Impact: Effects of the microenvironment on hiPSC-PCMs via costamere mechanotransduction may provide insights for engineering biopacemakers with a suitable ECM, to potentially preserve automaticity in hiPSC-PCMs and sustain long-term pacemaking function, making biopacemakers a step closer to reality. |
| format | Article |
| id | doaj-art-43e2727a2eee40e78dd75fd48aaa3038 |
| institution | Kabale University |
| issn | 2059-8661 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Cambridge University Press |
| record_format | Article |
| series | Journal of Clinical and Translational Science |
| spelling | doaj-art-43e2727a2eee40e78dd75fd48aaa30382025-08-20T03:40:18ZengCambridge University PressJournal of Clinical and Translational Science2059-86612025-04-01914114210.1017/cts.2024.1070479 Effects of extracellular matrix on pacemaking cardiomyocyte functionBrian Howard0Regan Smithers1Kaitlin Van Brusselen2Hillary K.J. Kao3Deborah K. Lieu4University of California, Davis Department of Internal Medicine, Division of Cardiovascular Medicine and Institute for Regenerative CuresUniversity of California, Davis Department of Internal Medicine, Division of Cardiovascular Medicine and Institute for Regenerative CuresUniversity of California, Davis Department of Internal Medicine, Division of Cardiovascular Medicine and Institute for Regenerative CuresUniversity of California, Davis Department of Internal Medicine, Division of Cardiovascular Medicine and Institute for Regenerative CuresUniversity of California, Davis Department of Internal Medicine, Division of Cardiovascular Medicine and Institute for Regenerative CuresObjectives/Goals: The extracellular matrix (ECM) of the sinoatrial node (SAN) is critical for maintaining automaticity in hiPSC-derived pacemaking cardiomyocytes (PCMs) under cyclic strain. We aim to determine the ECM ligands responsible for cell-ECM mediated mechanotransduction and the resulting phenotype in PCMs. Methods/Study Population: HiPSCs are differentiated to PCM and replated on substrate with 5 or 15 kPa PDMS that are coated with 5 or 25 ug/cm of either collagen I or fibronectin at sub-confluent density to restrict junction engagement to only costameres. Then, PCM are subjected to 10% cyclic mechanical strain at 1 Hz for 48 hours, with static culture as control. PCMs from all conditions are subsequently fixed and stained for cardiomyocyte-specific troponin T (TnT), pacemaking HCN4 channel, and pro-pacemaking transcription factors (Shox2, Isl1, Tbx3, Tbx18). Additionally, PCM cell size will also be assessed. Results/Anticipated Results: Considering the amount of hypertrophy and myofilament in CMs correlates with mechanical strain, we expect a reduced degree of mechanotransduction in hiPSC-PCM on collagen I with a stiffness 15 kPa to induce smaller cell size with fewer myofilament and an upregulation of HCN4 and pro-pacemaking transcription factors than those on 5 kPa and those on fibronectin of either 5 or 15 kPa after cyclic strain. This is because COL1 is reported to have a lower signaling threshold but a limited sensitivity to force which contributes to the diminished mechanotransduction signaling. Discussion/Significance of Impact: Effects of the microenvironment on hiPSC-PCMs via costamere mechanotransduction may provide insights for engineering biopacemakers with a suitable ECM, to potentially preserve automaticity in hiPSC-PCMs and sustain long-term pacemaking function, making biopacemakers a step closer to reality.https://www.cambridge.org/core/product/identifier/S2059866124010707/type/journal_article |
| spellingShingle | Brian Howard Regan Smithers Kaitlin Van Brusselen Hillary K.J. Kao Deborah K. Lieu 479 Effects of extracellular matrix on pacemaking cardiomyocyte function Journal of Clinical and Translational Science |
| title | 479 Effects of extracellular matrix on pacemaking cardiomyocyte function |
| title_full | 479 Effects of extracellular matrix on pacemaking cardiomyocyte function |
| title_fullStr | 479 Effects of extracellular matrix on pacemaking cardiomyocyte function |
| title_full_unstemmed | 479 Effects of extracellular matrix on pacemaking cardiomyocyte function |
| title_short | 479 Effects of extracellular matrix on pacemaking cardiomyocyte function |
| title_sort | 479 effects of extracellular matrix on pacemaking cardiomyocyte function |
| url | https://www.cambridge.org/core/product/identifier/S2059866124010707/type/journal_article |
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