Hiller Benjamin M, Marmion David J, Thompson Cayla A, Elliott Nathaniel A, Federoff Howard, Brundin Patrik, Mattis Virginia B, McMahon Christopher W, Kordower Jeffrey H
Department of Neurological Sciences, Rush University Medical Center, Chicago, IL, USA.
Department of Neurobiology, Barrow Neurological Institute, Phoenix, AZ, USA.
NPJ Regen Med. 2022 Apr 21;7(1):24. doi: 10.1038/s41536-022-00221-y.
In pursuit of treating Parkinson's disease with cell replacement therapy, differentiated induced pluripotent stem cells (iPSC) are an ideal source of midbrain dopaminergic (mDA) cells. We previously established a protocol for differentiating iPSC-derived post-mitotic mDA neurons capable of reversing 6-hydroxydopamine-induced hemiparkinsonism in rats. In the present study, we transitioned the iPSC starting material and defined an adapted differentiation protocol for further translation into a clinical cell transplantation therapy. We examined the effects of cellular maturity on survival and efficacy of the transplants by engrafting mDA progenitors (cryopreserved at 17 days of differentiation, D17), immature neurons (D24), and post-mitotic neurons (D37) into immunocompromised hemiparkinsonian rats. We found that D17 progenitors were markedly superior to immature D24 or mature D37 neurons in terms of survival, fiber outgrowth and effects on motor deficits. Intranigral engraftment to the ventral midbrain demonstrated that D17 cells had a greater capacity than D24 cells to innervate over long distance to forebrain structures, including the striatum. When D17 cells were assessed across a wide dose range (7,500-450,000 injected cells per striatum), there was a clear dose response with regards to numbers of surviving neurons, innervation, and functional recovery. Importantly, although these grafts were derived from iPSCs, we did not observe teratoma formation or significant outgrowth of other cells in any animal. These data support the concept that human iPSC-derived D17 mDA progenitors are suitable for clinical development with the aim of transplantation trials in patients with Parkinson's disease.
为了通过细胞替代疗法治疗帕金森病，分化的诱导多能干细胞（iPSC）是中脑多巴胺能（mDA）细胞的理想来源。我们之前建立了一种方案，可将iPSC分化为有丝分裂后mDA神经元，这些神经元能够逆转大鼠6-羟基多巴胺诱导的偏侧帕金森病。在本研究中，我们更换了iPSC起始材料，并确定了一种适应性分化方案，以便进一步转化为临床细胞移植疗法。我们通过将mDA祖细胞（在分化第17天，即D17时冻存）、未成熟神经元（D24）和有丝分裂后神经元（D37）移植到免疫受损的偏侧帕金森病大鼠体内，研究了细胞成熟度对移植细胞存活和疗效的影响。我们发现，在存活、纤维生长以及对运动缺陷的影响方面，D17祖细胞明显优于未成熟的D24或成熟的D37神经元。向腹侧中脑黑质内移植表明，D17细胞比D24细胞具有更强的远距离支配前脑结构（包括纹状体）的能力。当在较宽剂量范围内（每纹状体注射7500 - 450000个细胞）评估D17细胞时，在存活神经元数量、神经支配和功能恢复方面存在明显的剂量反应关系。重要的是，尽管这些移植物来源于iPSC，但我们在任何动物中均未观察到畸胎瘤形成或其他细胞的显著生长。这些数据支持了这样一种概念，即源自人类iPSC的D17 mDA祖细胞适合用于临床开发，旨在对帕金森病患者进行移植试验。
