Applying induced pluripotent stem cell derived endothelial cells to regenerative therapy in chronic limb threatening ischemia

外周动脉疾病(PAD)在退伍军人中非常普遍。退伍军人 与非退伍军人相比，PAD发病年龄更早，风险更高 经历外周动脉疾病的不良后果。慢性肢体威胁性缺血 外周动脉疾病(CLTI)是最严重的外周动脉疾病，是发病率的重要原因，包括 持续的休息疼痛，神经元功能障碍，最终失去肢体。而血管内和外科手术 治疗方案已经进步，许多患者由于以下原因不适合进行血运重建 解剖学上的限制或并存。对这些“无选择的CLTI”患者的治疗仅限于 由疼痛控制和大范围截肢组成的姑息疗法，这两者都会导致显著的发病率 和残疾。改进的治疗方式来治疗这一“非选择性CLTI”人群，并 防止肢体丧失是迫切需要的。 干细胞疗法已成为治疗这种“非选择性慢性感染”的一种很有前途的新方法。 人口。诱导多能干细胞(IPSC)技术的出现允许无限地 可复制和可扩展的干细胞来源。这项技术已被应用于血管 通过建立所述干细胞向无数血管的可靠的定向分化而引起的疾病 功能正常的血管所需的细胞类型。我们先前的工作克服了过早丢失的障碍 EC的功能和文化中的复制衰老，为下一步的发展打开了大门 IPSC-EC治疗学。 该方案旨在(1)验证临床级别的IPSC-ECs在植入体中的存活和功能 和可吸收的血小板裂解水凝胶，以及(2)量化肢体血流和功能的恢复 将含有IPSC-ECs的水凝胶植入缺血小鼠的后肢。这项工作将 建立关键的临床前动物模型，利用PAD患者来源的IPSC-ECs促进局部 肢体缺血血管再生作为治疗慢性盆腔感染的新方法。

Peripheral arterial disease (PAD) is highly prevalent among the Veteran population. Veterans experience PAD onset at an earlier age than their non-Veteran counterparts and are at increased risk of experiencing adverse outcomes from peripheral arterial disease. Chronic limb threatening ischemia (CLTI), the most severe form of peripheral arterial disease, is a significant cause of morbidity, including persistent pain at rest, neuronal dysfunction, and ultimately limb loss. While endovascular and surgical treatment options have advanced, many patients are not candidates for revascularization due to anatomic constraints or co-morbidities. Therapies for these “no-option CLTI” patients are limited to palliation consisting of pain control and major amputation, both of which lead to significant morbidity and disability. Improved therapeutic modalities to treat this “no-option CLTI” population and to prevent limb loss are desperately needed. Stem cell-based therapies have emerged as a promising new modality to treat this “no-option CLTI” population. The advent of induced pluripotent stem cell (iPSC) technology allows for an infinitely reproducible and scalable source of stem cells. This technology was rendered applicable to vascular disease by the establishment of reliable directed differentiation of said stem cells to the myriad vascular cell types required for a functional blood vessel. Our prior work overcame the barrier of premature loss of EC function and replicative senescence in culture, opening the door to the next steps in developing iPSC-EC therapeutics. This proposal aims to (1) validate the survival and function of clinical grade iPSC-ECs in implantable and resorbable platelet lysate hydrogels, and (2) quantify the restoration of limb perfusion and function after implantation of hydrogels containing iPSC-ECs into ischemic mouse hindlimbs. This work will generate a critical pre-clinical animal model of utilizing PAD patient-derived iPSC-ECs to promote local vascular regeneration in ischemic limbs as a novel therapy for CLTI.