Transforming growth factor-β1/Thrombospondin-1/CD47 axis mediates dysfunction in CD34+ cells derived from diabetic older adults

The role of telomerase reverse transcriptase in the mitochondrial protective functions of Angiotensin-(1–7) in diabetic CD34+ cells

“…This study is a logistical extension of our ongoing research studies focused at determining the role of TERT in the vascular-repair relevant functions in diabetic CD34 + cells that were stimulated by two different approaches, MasR activation by Ang-(1–7)[ 42 ] or TGFβ1-silencing. [ 46 ] PMO approach delivers the antisense to the extra and intranuclear compartments in a unique way of rapid delivery, which was used for accomplishing TGFβ1-silencing. [ 46 , 51 , 52 ] As hypothesized, protective functions of both approaches were found to be mitoTERT-dependent.…”

“…[ 46 ] PMO approach delivers the antisense to the extra and intranuclear compartments in a unique way of rapid delivery, which was used for accomplishing TGFβ1-silencing. [ 46 , 51 , 52 ] As hypothesized, protective functions of both approaches were found to be mitoTERT-dependent. Increasing mitoTERT by decreasing TERT-splicing is a novel mechanism of mitochondrial protection thereby restoring cellular functions in diabetes.…”

“…Silencing of TGF-β1 or blockade of β-TERT splicing was accomplished by using Phosphorodiamidate Morpholino Oligomer (PMO). A control (5′-CGTTCTGATAGCTGTACCTC-3′)-, TGFβ1 (5′-GCACTGCCGAGAGGGCGAACA-3′)- [ 25 , 46 ] and β-TERT-splicing blocking (5′-GGTCTGTCAAGGTAGTGAC-3′)-PMOs were obtained from Gene Tools (Philomath, OR) in lyophilized form and were dissolved in sterile distilled water. PMO approach delivers the antisense to the extra and intranuclear compartments in a unique way of rapid delivery and the time-course of entry and efflux of PMOs in CD34 + cells was reported previously.…”

“…PMO approach delivers the antisense to the extra and intranuclear compartments in a unique way of rapid delivery and the time-course of entry and efflux of PMOs in CD34 + cells was reported previously. [ 46 , 51 , 52 ] CD34 + cells were incubated at 6 μg/ml for control- or TGFβ1-PMO in StemSpan for 16–18 h at 37 °C, and then excess PMO was removed by centrifugation with PBS as described above. Then, the cells were either used for in vitro assays or preserved at −80°C.…”

“…[ 45 ] Interestingly, TGFβ1-silencing also showed similar protective functions as that of Ang-(1–7) by restoring NO generation, proliferation and migration to areas of ischemic injury. [ 46 , 47 ] While no reports showed evidence for the involvement of TERT in the protective functions of Ang-(1–7) in CD34 + cells, in coronary arteries Ang-(1–7) increased TERT expression and promoted mitoTERT-dependent NO-mediated relaxation. [ 48 , 49 ] These studies led to the hypothesis that the protective functions of Ang-(1–7) or TGFβ1-silencing in diabetic CD34 + cells are mediated by mitoTERT.…”