Table_1_ALDH1A3 Is the Key Isoform That Contributes to Aldehyde Dehydrogenase Activity and Affects in Vitro Proliferation in Cardiac Atrial Appendage Progenitor Cells.DOCX

<p>High aldehyde dehydrogenase (ALDH^hi) activity has been reported in normal and cancer stem cells. We and others have shown previously that human ALDH^hi cardiac atrial appendage cells are enriched with stem/progenitor cells. The role of ALDH in these cells is poorly understood but it may come down to the specific ALDH isoform(s) expressed. This study aimed to compare ALDH^hi and ALDH^lo atrial cells and to identify the isoform(s) that contribute to ALDH activity, and their functional role.</p><p>Methods and Results: Cells were isolated from atrial appendage specimens from patients with ischemic and/or valvular heart disease undergoing heart surgery. ALDH^hi activity assessed with the Aldefluor reagent coincided with primitive surface marker expression (CD34⁺). Depending on their ALDH activity, RT-PCR analysis of ALDH^hi and ALDH^lo cells demonstrated a differential pattern of pluripotency genes (Oct 4, Nanog) and genes for more established cardiac lineages (Nkx2.5, Tbx5, Mef2c, GATA4). ALDH^hi cells, but not ALDH^lo cells, formed clones and were culture-expanded. When cultured under cardiac differentiation conditions, ALDH^hi cells gave rise to a higher number of cardiomyocytes compared with ALDH^lo cells. Among 19 ALDH isoforms known in human, ALDH1A3 was most highly expressed in ALDH^hi atrial cells. Knocking down ALDH1A3, but not ALDH1A1, ALDH1A2, ALDH2, ALDH4A1, or ALDH8A1 using siRNA decreased ALDH activity and cell proliferation in ALDH^hi cells. Conversely, overexpressing ALDH1A3 with a retroviral vector increased proliferation in ALDH^lo cells.</p><p>Conclusions: ALDH1A3 is the key isoform responsible for ALDH activity in ALDH^hi atrial appendage cells, which have a propensity to differentiate into cardiomyocytes. ALDH1A3 affects in vitro proliferation of these cells.</p