Early-life peripheral infections reprogram retinal microglia and aggravate neovascular age-related macular degeneration in later life
Masayuki Hata, … , Ariel M. Wilson, Przemyslaw Sapieha
Masayuki Hata, … , Ariel M. Wilson, Przemyslaw Sapieha
Published February 15, 2023
Citation Information: J Clin Invest. 2023;133(4):e159757. https://doi.org/10.1172/JCI159757.
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Research Article Ophthalmology

Early-life peripheral infections reprogram retinal microglia and aggravate neovascular age-related macular degeneration in later life

Abstract

Pathological neovascularization in age-related macular degeneration (nvAMD) drives the principal cause of blindness in the elderly. While there is a robust genetic association between genes of innate immunity and AMD, genome-to-phenome relationships are low, suggesting a critical contribution of environmental triggers of disease. Possible insight comes from the observation that a past history of infection with pathogens such as Chlamydia pneumoniae, or other systemic inflammation, can predispose to nvAMD in later life. Using a mouse model of nvAMD with prior C. pneumoniae infection, endotoxin exposure, and genetic ablation of distinct immune cell populations, we demonstrated that peripheral infections elicited epigenetic reprogramming that led to a persistent memory state in retinal CX3CR1+ mononuclear phagocytes (MNPs). The immune imprinting persisted long after the initial inflammation had subsided and ultimately exacerbated choroidal neovascularization in a model of nvAMD. Single-cell assay for transposase-accessible chromatin sequencing (scATAC-seq) identified activating transcription factor 3 (ATF3) as a central mediator of retina-resident MNP reprogramming following peripheral inflammation. ATF3 polarized MNPs toward a reparative phenotype biased toward production of proangiogenic factors in response to subsequent injury. Therefore, a past history of bacterial endotoxin–induced inflammation can lead to immunological reprograming within CNS-resident MNPs and aggravate pathological angiogenesis in the aging retina.

Authors

Masayuki Hata, Maki Hata, Elisabeth M.M.A. Andriessen, Rachel Juneau, Frédérique Pilon, Sergio Crespo-Garcia, Roberto Diaz-Marin, Vera Guber, Francois Binet, Frédérik Fournier, Manuel Buscarlet, Caroline Grou, Virginie Calderon, Emilie Heckel, Heather J. Melichar, Jean-Sebastien Joyal, Ariel M. Wilson, Przemyslaw Sapieha

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Figure 9

Peripheral exposure to endotoxins modulates myeloid cell response via ATF3 deregulation.

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Peripheral exposure to endotoxins modulates myeloid cell response via AT...
(A) Volcano plot of accessible regions with differentially accessible regions (DARs; defined by an FDR adjusted P value < 0.05, total of 51 DARs) identified between comparisons of microglial 4×LPS cluster (C3) versus microglial PBS cluster (C2) as found by CX3CR1+ retinal myeloid cell scATAC-seq data. (B) Cis-coaccessibility network (CCAN) links between the ATF3 promoter and distal sites in the surrounding region generated by subset analysis by cluster. Connections from microglial PBS cluster (C2) and microglial 4×LPS cluster (C3) are shown, with a minimum coaccessibility score of 0.3. (C) Chromatin accessibility in the promoter region of Atf3 in BMDMs from PBS-pretreated and 4×LPS-pretreated mice as analyzed by qPCR. n = 4 (PBS), n = 4 (4×LPS). (D) Representative immunoblots showing p-NF-κB, total NF-κB, p-c-JUN, total c-JUN, and ATF3 expression in BMDMs from PBS-pretreated and 4×LPS-pretreated mice with and without LPS restimulation. (E) Schematic representation of knockdown of Atf3 gene expression in BMDMs using siRNA. C57BL/6J mice were treated with 4×LPS or PBS at 7 weeks of age and BM cells were collected at 11 weeks of age. BM cells were differentiated into BMDMs with M-CSF. BMDMs were transfected with Atf3 or control siRNA for 24 hours and then restimulated with LPS for 4 hours. RNA was extracted for qPCR analysis. (F–J) mRNA expression in BMDMs transfected with Atf3 or control siRNA from PBS-pretreated and 4×LPS-pretreated mice with LPS restimulation: Atf3 (F), Tnf (G), Il6 (H), Vegfa (I), and Pdgfb (J); n = 6 for all groups. Data are presented as mean ± SEM. Comparisons between groups were analyzed using Student’s unpaired t test (C) or 1-way ANOVA with Tukey’s multiple-comparison test (F–J). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.