Calzi Sergio Li, Chakraborty Dibyendu, Hu Ping, Prasad Ram, Adu-Rutledge Yvonne, Vieira Cristiano, Sheini Fadeela, Boulton Michael E, Yoder Mervin C, Cheng Changde, Grant Maria B
Department of Ophthalmology and Visual Sciences, University of Alabama at Birmingham (UAB), Birmingham, AL 35294, USA.
Schepens Eye Research Institute, Harvard Medical School, and Massachusetts General Hospital, Boston, MA 02114, USA.
Cells. 2025 Aug 30;14(17):1352. doi: 10.3390/cells14171352.
To investigate the therapeutic potential of inducible pluripotent stem cell (hiPSC)-based vascular repair, we evaluated two vascular reparative cell populations, CD34 cells derived from hiPSC (hiPSC-CD34) and endothelial colony forming cells (ECFCs) derived from hiPSC (iPS-ECFCs), alone and in combination, in a type 2 diabetic (db/db) mouse model of DR. hiPSC-CD34 cells (1 × 10) or iPSC- ECFCs (1 × 10) alone or in combination (1.1 × 10) were injected into the vitreous of immunosuppressed db/db mice with six months of established diabetes. One month post-injection, mice underwent electroretinography (ERG) and optical coherence tomography (OCT) to evaluate functional and structural retinal recovery with iPSC administration. Immunohistochemistry (IHC) was used to assess recruitment and incorporation of cells into the retinal vasculature. Retinas from the experimental groups were analyzed using Functional Proteomics via Reverse Phase Protein Array (RPPA). Functional assessment via ERG demonstrated significant improvements in retinal response in the diabetic cohorts treated with either hiPSC-derived CD34 cells or hiPSC-ECFCs. Retinal thickness, assessed by OCT, was restored to near-nondiabetic levels in mice treated with hiPSC-CD34 cells alone and the combination group, whereas hiPSC-ECFCs alone did not significantly affect retinal thickness. One month following intravitreal injection, hiPSC-CD34 cells were localized to perivascular regions, whereas hiPSC-ECFCs were observed to integrate directly into the retinal vasculature. RPPA analysis revealed interaction-significant changes, and this was interpreted as a combination-specific, non-additive host responses (mA, PI3K-AKT-mTOR, glycolysis, endothelial junction pathways). The studies support that injection of hiPSC-CD34 cells and hiPSC-ECFCs, both individually and in combination, showed benefit; however, iPSC combination-specific effects were identified by measurement of retinal thickness and by RPPA.
为了研究基于诱导多能干细胞（hiPSC）的血管修复的治疗潜力，我们在糖尿病视网膜病变（DR）的2型糖尿病（db/db）小鼠模型中，单独或联合评估了两种血管修复细胞群，即源自hiPSC的CD34细胞（hiPSC-CD34）和源自hiPSC的内皮集落形成细胞（iPS-ECFCs）。将hiPSC-CD34细胞（1×10）或iPS-ECFCs（1×10）单独或联合（1.1×10）注射到患有6个月已确诊糖尿病的免疫抑制db/db小鼠的玻璃体中。注射后1个月，对小鼠进行视网膜电图（ERG）和光学相干断层扫描（OCT），以评估iPSC给药后视网膜功能和结构的恢复情况。免疫组织化学（IHC）用于评估细胞募集以及细胞整合到视网膜血管系统中的情况。通过反相蛋白质阵列（RPPA）使用功能蛋白质组学对实验组的视网膜进行分析。通过ERG进行的功能评估表明，用源自hiPSC的CD34细胞或hiPSC-ECFCs治疗的糖尿病组视网膜反应有显著改善。通过OCT评估，单独用hiPSC-CD34细胞治疗的小鼠和联合治疗组的视网膜厚度恢复到接近非糖尿病水平，而单独使用hiPSC-ECFCs对视网膜厚度没有显著影响。玻璃体内注射1个月后，hiPSC-CD34细胞定位于血管周围区域，而hiPSC-ECFCs则直接整合到视网膜血管系统中。RPPA分析揭示了相互作用的显著变化，这被解释为组合特异性、非加性的宿主反应（mA、PI3K-AKT-mTOR、糖酵解、内皮连接途径）。这些研究支持，单独或联合注射hiPSC-CD34细胞和hiPSC-ECFCs均显示出益处；然而，通过测量视网膜厚度和RPPA确定了iPSC组合特异性效应。
