Redirected T Cell Therapy to Cure Invasive Fungal Infections

项目摘要 嵌合抗原受体(CAR)T细胞治疗给耐药感染患者带来新希望 曲霉、念珠菌或毛霉等生物体。这是第一次模式识别受体 (Dectin-1)已被用于重定向T细胞特异性以控制真菌感染。DECIN-1轿车(D-CAR)可以 激活细胞溶解机制，以及可能的穿孔素/颗粒酶和颗粒溶素途径 T细胞。从D-CAR T细胞产生干扰素-可能进一步增强对侵袭性真菌的天然免疫 如果重组干扰素-γ是通过药物途径给药，或来源于CD4h辅助T细胞或天然来源，则感染 杀手细胞。 在R21阶段，限制CAR T细胞疗法立即临床应用的两个主要因素将是 解决：(1)快速增殖的β-葡聚糖特异性D-CAR T细胞的产生和(2)体内长期 坚持控制侵袭性真菌感染。 目前有几种类型的CARS被用于临床试验，以控制B细胞恶性肿瘤。因为现在还不是 显然，哪种汽车设计为给定的患者提供了完全合格的T细胞激活，我们已经开发出一种 筛选多个CAR分子的方法。我们团队已经开发了EZ-CAR平台，用于发电 通过混合和匹配来自已知T细胞激活受体的成分来实现多个CARS，同时保持 目标区域完好无损。使用这种方法，我们将产生大约21辆带有Dectin-1真菌的D-Cars 目标域。快速生产(从捐献者采集PBMC后10天内)可能会改善治疗效果 人工制造的T细胞的潜力，因为它避免了复制介导的T细胞衰老和终末 与体内持久性丧失相关的分化。 在R33阶段，这项研究将扩大到针对各种临床上重要的机会性霉菌。 (毛霉、生孢霉)和酵母菌(念珠菌)。MD Anderson临床分离株耐药菌株的鉴定 实验室将用于验证D-CAR T细胞的治疗效果。在一些真菌中，例如 根霉(毛霉目)，β-葡聚糖层被糖胺聚糖(GAG)层掩盖。D-CAR T- 细胞疗法将与卡泊芬净等真菌细胞壁生物合成抑制剂结合使用，以破坏 糖胺多糖层，这将使D-CAR T细胞疗法得到更好的识别和激活。 综上所述，患有侵袭性真菌感染的患者由于原发免疫缺陷，如基因 突变和继发性免疫缺陷，如人类免疫缺陷病毒感染、癌症和 移植极有可能从免疫辅助治疗中受益。单工程T-型发动机的研制 可针对各种病原体的细胞，如D-CAR T细胞，将T细胞特异性重定向至 曲霉菌、念珠菌和毛霉菌是非常有必要对抗侵袭性真菌感染的 免疫功能受损的病人。

Project Summary Chimeric antigen receptor (CAR) T-cell therapy give new hope to patients suffering from drug-resistant infectious organisms such as Aspergillus, Candida, or Mucor. This is the first time that a pattern-recognition receptor (Dectin-1) has been adapted to redirect T-cell specificity to control fungal infection. Dectin-1 CAR (D-CAR) can activate the cytolytic machinery, and likely the perforin/granzyme and granulysin pathway, of genetically modified T-cells. The production of IFN- from the D-CAR+ T-cells may further augment innate immunity to invasive fungal infections if recombinant IFN-γ is administered pharmacologically or derived from CD4+ helper T-cells or natural killer cells. In the R21 phase, 2 major factors that limit immediate clinical applications of CAR T-cell therapy will be addressed: (1) generation of rapidly proliferative β-glucan-specific D-CAR+ T-cells and (2) long-term in vivo persistence to control invasive fungal infection. Several types of CARs are currently used in clinical trials to control B-cell malignancy. Because it is not yet apparent which CAR design provides fully competent T-cell activation for a given patient, we have developed an approach for screening multiple CAR molecules. Our team has developed the EZ-CAR platform for generating multiple CARs by mixing and matching components derived from known T-cell activating receptors while keeping the targeting domain intact. Using this approach, we will generate about 21 D-CARs with the Dectin-1 fungal targeting domain. Rapid production (within 10 days of PBMC collection from donor) may improve the therapeutic potential of the manufactured T-cells because it avoids the replication-mediated T-cell senescence and terminal differentiation that is associated with loss of in vivo persistence. In the R33 phase, the study will be expanded to target a wide variety of clinically important opportunistic molds (Mucor, Scedosporium) and yeasts (Candida). Drug-resistant isolates identified in MD Anderson clinical laboratories will be used for validating the therapeutic efficacy of the D-CAR+ T cells. In some fungi, such as Rhizopus (Mucorales family), the β-glucan layer is masked by the glycosaminoglycans (GAG) layer. D-CAR+ T- cell therapy will be used in combination with fungal cell wall biosynthesis inhibitors such as caspofungin to disrupt the glycosaminoglycans layer, which will allow better recognition and activation of the D-CAR+ T-cell therapy. In summary, patients suffering from invasive fungal infections due to primary immunodeficiencies such as genetic mutations and secondary immunodeficiencies such as human immunodeficiency virus infection, cancer, and transplantation are highly likely to benefit from immune adjuvant therapy. Development of single-engineered T- cells that can target various pathogens, such as D-CAR+ T-cells cells, which redirect T-cell specificity to Aspergillus, Candida, and Mucor species, is highly warranted to combat invasive fungal infections in immunocompromised patients.