Tang Yinghua, Fakhari Shahab, Huntemann Eric D, Feng Zehua, Wu Peipei, Feng William Y, Lei Junying, Yuan Feng, Excoffon Katherine J, Wang Kai, Limberis Maria P, Kolbeck Roland, Yan Ziying, Engelhardt John F
Department of Anatomy & Cell Biology, University of Iowa Carver College of Medicine, Iowa City, IA 52242, USA.
Spirovant Science, Inc., Philadelphia, PA 19104, USA.
Mol Ther Methods Clin Dev. 2023 Mar 2;29:70-80. doi: 10.1016/j.omtm.2023.02.015. eCollection 2023 Jun 8.
The efficacy of redosing the recombinant adeno-associated virus (rAAV) vector rAAV2.5T to ferret lung is limited by AAV neutralizing antibody (NAb) responses. While immunosuppression strategies have allowed for systemic rAAV repeat dosing, their utility for rAAV lung-directed gene therapy is largely unexplored. To this end, we evaluated two immunosuppression (IS) strategies to improve repeat dosing of rAAV2.5T to ferret lungs: (1) a combination of three IS drugs (Tri-IS) with broad coverage against cellular and humoral responses (methylprednisolone [MP], azathioprine, and cyclosporine) and (2) MP alone, which is typically used in systemic rAAV applications. Repeat dosing utilized AAV2.5T-SP183-fCFTRΔR (recombinant ferret transgene), followed 28 days later by AAV2.5T-SP183-gLuc (for quantification of transgene expression). Both the Tri-IS and MP strategies significantly improved transgene expression following repeat dosing and reduced AAV2.5T NAb responses in the bronchioalveolar lavage fluid (BALF) and plasma, while AAV2.5T binding antibody subtypes and cellular immune responses by ELISpot were largely unchanged by IS. One exception was the reduction in plasma AAV2.5T binding immunoglobulin G (IgG) in both IS groups. Only the Tri-IS strategy significantly suppressed splenocyte expression of (interferon α [IFN-α]) and Our studies suggest that IS strategies may be useful in clinical application of rAAV targeting lung genetic diseases such as cystic fibrosis.
重组腺相关病毒（rAAV）载体rAAV2.5T再次给药至雪貂肺部的疗效受到AAV中和抗体（NAb）反应的限制。虽然免疫抑制策略已允许全身性rAAV重复给药，但其在rAAV肺靶向基因治疗中的效用在很大程度上尚未得到探索。为此，我们评估了两种免疫抑制（IS）策略，以改善rAAV2.5T再次给药至雪貂肺部的情况：（1）三种具有广泛细胞和体液反应覆盖范围的IS药物组合（三联IS）（甲基强的松龙[MP]、硫唑嘌呤和环孢素），以及（2）单独使用MP，这通常用于全身性rAAV应用。重复给药使用AAV2.5T-SP183-fCFTRΔR（重组雪貂转基因），28天后再使用AAV2.5T-SP183-gLuc（用于转基因表达定量）。三联IS和MP策略均显著改善了重复给药后的转基因表达，并降低了支气管肺泡灌洗液（BALF）和血浆中AAV2.5T NAb反应，而IS对AAV2.5T结合抗体亚型和ELISpot检测的细胞免疫反应基本无影响。一个例外是两个IS组血浆中AAV2.5T结合免疫球蛋白G（IgG）均减少。只有三联IS策略显著抑制了脾细胞中（干扰素α[IFN-α]）和的表达。我们的研究表明，IS策略可能在rAAV靶向肺遗传疾病如囊性纤维化的临床应用中有用。
