Panjwani M Kazim, Smith Jenessa B, Schutsky Keith, Gnanandarajah Josephine, O'Connor Colleen M, Powell Daniel J, Mason Nicola J
Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.
Mol Ther. 2016 Sep;24(9):1602-14. doi: 10.1038/mt.2016.146. Epub 2016 Jul 12.
Preclinical murine models of chimeric antigen receptor (CAR) T cell therapy are widely applied, but are greatly limited by their inability to model the complex human tumor microenvironment and adequately predict safety and efficacy in patients. We therefore sought to develop a system that would enable us to evaluate CAR T cell therapies in dogs with spontaneous cancers. We developed an expansion methodology that yields large numbers of canine T cells from normal or lymphoma-diseased dogs. mRNA electroporation was utilized to express a first-generation canine CD20-specific CAR in expanded T cells. The canine CD20 (cCD20) CAR expression was efficient and transient, and electroporated T cells exhibited antigen-specific interferon-gamma (IFN-γ) secretion and lysed cCD20+ targets. In a first-in-canine study, autologous cCD20-ζ CAR T cells were administered to a dog with relapsed B cell lymphoma. Treatment was well tolerated and led to a modest, but transient, antitumor activity, suggesting that stable CAR expression will be necessary for durable clinical remissions. Our study establishes the methodologies necessary to evaluate CAR T cell therapy in dogs with spontaneous malignancies and lays the foundation for use of outbred canine cancer patients to evaluate the safety and efficacy of next-generation CAR therapies and their optimization prior to translation into humans.
嵌合抗原受体（CAR）T细胞疗法的临床前小鼠模型被广泛应用，但由于无法模拟复杂的人类肿瘤微环境以及充分预测对患者的安全性和疗效而受到极大限制。因此，我们试图开发一种系统，使我们能够在患有自发性癌症的犬类中评估CAR T细胞疗法。我们开发了一种扩增方法，可从正常或患有淋巴瘤的犬类中产生大量犬T细胞。利用mRNA电穿孔在扩增的T细胞中表达第一代犬CD20特异性CAR。犬CD20（cCD20）CAR表达高效且短暂，电穿孔的T细胞表现出抗原特异性干扰素-γ（IFN-γ）分泌并裂解cCD20+靶标。在一项犬类首例研究中，将自体cCD20-ζ CAR T细胞施用于一只复发性B细胞淋巴瘤犬。治疗耐受性良好，并导致适度但短暂的抗肿瘤活性，这表明稳定的CAR表达对于持久的临床缓解是必要的。我们的研究确立了在患有自发性恶性肿瘤的犬类中评估CAR T细胞疗法所需的方法，并为使用远交系犬类癌症患者评估下一代CAR疗法的安全性和疗效以及在转化为人类之前对其进行优化奠定了基础。
