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. 2020 Aug 6;11:3929. doi: 10.1038/s41467-020-17577-8

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 9 — a Study design to assess survival following AAV-hSPB plus BLES gene therapy in inherited SP-B deficient adult mice. Control mice underwent sham surgeries with IT injections of 1× PBS plus BLES 1 day (-D1) after dox removal (blue line). Treatment mice had dox removed 2 days (-D2; dashed pink line) and 1 day (-D1; solid pink line) before IT administration of 10¹¹ vg per mouse of AAV-hSPB plus BLES. b Kaplan-Meier survival curves from IT injections of a 10¹¹ vg dose of AAV-hSPB plus BLES 1 (-D1) and 2 (-D2; ***P = 0.0001) days after dox removal. c Mean weight measurements with SD of all surviving mice from b. d Study design to assess survival following IT delivery of 10¹¹ vg per mouse AAV-hSPB plus BLES into P13 to P15 neonatal SP-B deficient mice. The pups were removed from dox immediately following AAV administration by placing them with wild type FVB/N foster mothers that had recently given birth to similar aged pups. Two mice were euthanized from each group 3 days after surgery for histological lung examination. e Kaplan-Meier survival curves following IT delivery of 10¹¹ vg per mouse of AAV-hSPB plus BLES into P13 to P15 neonatal mice with dox removal immediately following surgery (***P = 0.0004 compared to 1× PBS). Neonatal pups were untreated and maintained on dox (black line), underwent sham surgeries with IT injections of 1× PBS (blue line; ***P = 0.0004 compared to on dox), or were IT administered 10¹¹ vg of AAV-hSPB plus BLES (pink line; **P = 0.0021 compared to on dox). The survival curve data do not include the mice removed for lung histology. f Mean weight measurements with SD of all surviving mice from e. g Gross lung images from all 3 groups (n = 2 per group) at harvest 3 days following AAV administration plus dox removal. Yellow arrows indicate regions of lung injury. h Representative H&E and WGJ staining of paraffin embedded whole right and left lungs 3 days after AAV administration and dox removal from all 3 groups in d (H&E and WGJ scale bar, 100 μm). (All survival curve P values in Fig. 9 = 2-tailed Log-rank, Mantel-Cox test with Bonferroni correction, significance is set at P = 0.0167, ns = not significant).