使用工程化 CD4+ 细胞归巢 mRNA-LNP 进行高效 CD4+ T 细胞靶向和基因重组,Molecular Therapy

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使用工程化 CD4+ 细胞归巢 mRNA-LNP 进行高效 CD4+ T 细胞靶向和基因重组
Molecular Therapy ( IF 12.1 ) Pub Date : 2021-06-04 , DOI: 10.1016/j.ymthe.2021.06.004
István Tombácz ₁ , Dorottya Laczkó ₁ , Hamna Shahnawaz ₁ , Hiromi Muramatsu ₁ , Ambika Natesan ₁ , Amir Yadegari ₁ , Tyler E Papp ₁ , Mohamad-Gabriel Alameh ₁ , Vladimir Shuvaev ₂ , Barbara L Mui ₃ , Ying K Tam ₃ , Vladimir Muzykantov ₂ , Norbert Pardi ₁ , Drew Weissman ₁ , Hamideh Parhiz ₁

Affiliation

核苷修饰的信使 RNA (mRNA)-脂质纳米粒子 (LNP) 是前两种 EUA（紧急使用授权）COVID-19 疫苗的基础。使用核苷修饰的 mRNA 作为药物制剂为治疗、预防和诊断分子干预开辟了巨大的机会。特别是，基于 mRNA 的药物可以特异性调节免疫细胞，例如 T 淋巴细胞，用于肿瘤、感染和其他疾病的免疫治疗。然而，关键的挑战是 T 细胞对外源 mRNA 的转染具有很强的抵抗力。在此，我们报告将 CD4 抗体与 LNP 结合，能够对 CD4+ 细胞（包括 T 细胞）进行特异性靶向和 mRNA 干预。小鼠全身注射后，CD4靶向放射性标记的mRNA-LNP在脾脏中积累，与非靶向mRNA-LNP相比，从脾脏分离的T细胞中提供的报告基因mRNA信号高出约30倍。静脉注射载有 Cre 重组酶编码 mRNA 的 CD4 靶向 LNP 提供了特异性剂量依赖性 loxP 介导的基因重组，导致脾脏和淋巴结中约 60% 和 40% 的 CD4+ T 细胞分别表达报告基因。 T 细胞表型分析显示 T 细胞亚群的转染一致，幼稚细胞、中枢记忆细胞和效应细胞中 CD4 靶向 mRNA-LNP 的摄取没有变化。本研究建立的mRNA特异性、高效靶向和转染至T细胞的技术为破坏性病症的免疫治疗和艾滋病毒治愈提供了平台技术。

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Highly efficient CD4+ T cell targeting and genetic recombination using engineered CD4+ cell-homing mRNA-LNPs

Nucleoside-modified messenger RNA (mRNA)-lipid nanoparticles (LNPs) are the basis for the first two EUA (Emergency Use Authorization) COVID-19 vaccines. The use of nucleoside-modified mRNA as a pharmacological agent opens immense opportunities for therapeutic, prophylactic and diagnostic molecular interventions. In particular, mRNA-based drugs may specifically modulate immune cells, such as T lymphocytes, for immunotherapy of oncologic, infectious and other conditions. The key challenge, however, is that T cells are notoriously resistant to transfection by exogenous mRNA. Here, we report that conjugating CD4 antibody to LNPs enables specific targeting and mRNA interventions to CD4+ cells, including T cells. After systemic injection in mice, CD4-targeted radiolabeled mRNA-LNPs accumulated in spleen, providing ∼30-fold higher signal of reporter mRNA in T cells isolated from spleen as compared with non-targeted mRNA-LNPs. Intravenous injection of CD4-targeted LNPs loaded with Cre recombinase-encoding mRNA provided specific dose-dependent loxP-mediated genetic recombination, resulting in reporter gene expression in about 60% and 40% of CD4+ T cells in spleen and lymph nodes, respectively. T cell phenotyping showed uniform transfection of T cell subpopulations, with no variability in uptake of CD4-targeted mRNA-LNPs in naive, central memory, and effector cells. The specific and efficient targeting and transfection of mRNA to T cells established in this study provides a platform technology for immunotherapy of devastating conditions and HIV cure.

更新日期：2021-06-04

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