Single-cell profiling of immune system alterations in lymphoid, barrier and solid tissues in aged mice

Krishnarajah, Sinduya; Ingelfinger, Florian; Friebel, Ekaterina; Cansever, Dilay; Amorim, Ana; Andreadou, Myrto; Bamert, David; Litscher, Gioana; Lutz, Mirjam; Mayoux, Maud; Mundt, Sarah; Ridder, Frederike; Sparano, Colin; Stifter, Sebastian A.; Ulutekin, Can; Unger, Susanne; Vermeer, Marijne; Zwicky, Pascale; Greter, Melanie; Tugues, Sònia; Feo, Donatella De

doi:10.1038/s43587-021-00148-x

Cited by 21 publications

(14 citation statements)

References 81 publications

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“…We therefore hypothesized that if repeated rupture events accumulate over time in vivo, WT AM should gradually acquire a signature related to Lamin A/C CKO signature. In mice aged for 63 weeks, we detected a reduction of AM (Figure S8A), in agreement with a drop in AM levels previously reported in mice aged for more than 80 weeks (Krishnarajah et al, 2021;Wong et al, 2017;Xin et al, 2015). Next, we analyzed gene expression in AM from mice of increasing age using the Tabula Muris Senis (TMS) scRNAseq atlas, a resource characterizing aging in mouse tissues (The Tabula Muris Consortium et al, 2020).…”

Section: Lamin A/c Protects Against Acquisition Of Hallmarks Of Aging...supporting

confidence: 92%

Nuclear envelope disruption triggers hallmarks of aging in lung alveolar macrophages

Silva

Nader

Nadalin

et al. 2022

Preprint

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Aging is characterized by gradual immune dysfunction and increased risk for many diseases, including respiratory infections. Genomic instability is thought to play a central role in the aging process but the mechanisms that damage nuclear DNA in aging are insufficiently defined. Cells that migrate or reside within confined environments experience forces applied to their nucleus, leading to transient nuclear envelope (NE) ruptures. NE ruptures are associated with DNA damage, and Lamin A/C is required to limit these events. Here, we show that Lamin A/C protects lung alveolar macrophages from NE rupture and hallmarks of aging. Lamin A/C ablation in immune cells results in a selective depletion of lung alveolar macrophages (AM) and a heightened susceptibility to influenza infection. Lamin A/C-deficient AM that persist display constitutive nuclear envelope rupture marks, DNA damage and p53-dependent senescence. In wild-type mice, we found that AM migrate within constricted spaces in vivo, at heights that induce NE rupture and DNA damage. AM from aged wild-type mice and from Lamin A/C-deficient mice share an upregulated lysosomal signature with CD63 expression, and we find that CD63 is required to clear damaged DNA in macrophages. We propose that induction of genomic instability by NE disruption represents a mechanism of aging in alveolar macrophages.

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Section: Lamin A/c Protects Against Acquisition Of Hallmarks Of Aging...supporting

confidence: 92%

Nuclear envelope disruption triggers hallmarks of aging in lung alveolar macrophages

Silva

Nader

Nadalin

et al. 2022

Preprint

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“…Within this myeloid population, F4/80− cluster of differentiation 64 (CD64)− lymphocyte antigen 6C (Ly6C)+ cells were unchanged, F4/80+ CD64+ Ly6C+ cells were significantly increased, and F4/80+ CD64+ Ly6C− cells were significantly decreased ( p = 0.8154, 0.0050, 0.0029, respectively; Figure 6 A,B, Supplementary Figure S2 ). Since CD64 is expressed primarily on macrophages, and Ly6C is thought to be specific to circulation-derived inflammatory monocytes and macrophages, the increase in F4/80+ macrophages in Ltf iCre/+ Arid1a f/f uteri appears to be driven by infiltration of circulating Ly6C+ inflammatory cells rather than new growth of resident macrophages [ 33 , 34 , 35 , 36 , 37 , 38 ]. Further myeloid immune cell analysis revealed that Ly6G+ neutrophil numbers were consistent between genotypes, but CD64− cluster of differentiation 24 (CD24)+ uDCs, important for proper decidua formation, were significantly decreased in Ltf iCre/+ Arid1a f/f mice [ 13 , 38 ] ( p = 0.7818, 0.0016, respectively; Figure 6 C,D, Supplementary Figure S2 ).…”

Section: Resultsmentioning

confidence: 99%

“…Immunostaining identified a uterus-specific increase in total F4/80+ macrophages when endometrial epithelial Arid1a was deleted, but this analysis could not determine the source or characteristics of the cells since F4/80 marks tissue-resident macrophages as well as macrophages derived from circulating monocytes [ 38 ]. In our flow cytometry analysis, we were able to discriminate among F4/80+ CD64+ macrophages utilizing the monocyte and monocyte-derived cell marker Ly6C [ 33 , 35 , 37 , 38 ]. Since the Ly6C+ population increased in Ltf iCre/+ Arid1a f/f uteri and the Ly6C− population decreased, we conclude that the increased F4/80+ macrophages in the uterus were derived from circulating inflammatory monocytes [ 33 , 35 , 37 , 38 ].…”

Section: Discussionmentioning

confidence: 99%

“…In our flow cytometry analysis, we were able to discriminate among F4/80+ CD64+ macrophages utilizing the monocyte and monocyte-derived cell marker Ly6C [ 33 , 35 , 37 , 38 ]. Since the Ly6C+ population increased in Ltf iCre/+ Arid1a f/f uteri and the Ly6C− population decreased, we conclude that the increased F4/80+ macrophages in the uterus were derived from circulating inflammatory monocytes [ 33 , 35 , 37 , 38 ]. Since the F4/80− CD64− Ly6C+ cell populations were not different between genotypes, it appears that the altered cell type should be defined as an inflammatory monocyte-derived macrophage population rather than a classical monocyte population.…”

Section: Discussionmentioning

confidence: 99%

“…Since the F4/80− CD64− Ly6C+ cell populations were not different between genotypes, it appears that the altered cell type should be defined as an inflammatory monocyte-derived macrophage population rather than a classical monocyte population. However, we cannot conclude this with certainty since some monocytes also express F4/80 and CD64, and there is disagreement over whether Ly6C+ cells should be labeled as macrophages [ 33 , 35 , 37 , 38 ].…”

Section: Discussionmentioning

confidence: 99%

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Endometrial Epithelial ARID1A Is Required for Uterine Immune Homeostasis during Early Pregnancy

Marquardt

Ahn

Reske

et al. 2022

IJMS

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A growing body of work suggests epigenetic dysregulation contributes to endometriosis pathophysiology and female infertility. The chromatin remodeling complex subunit AT-rich interaction domain 1A (ARID1A) must be properly expressed to maintain normal uterine function. Endometrial epithelial ARID1A is indispensable for pregnancy establishment in mice through regulation of endometrial gland function; however, ARID1A expression is decreased in infertile women with endometriosis. We hypothesized that ARID1A performs critical operations in the endometrial epithelium necessary for fertility besides maintaining gland function. To identify alterations in uterine gene expression resulting from loss of epithelial ARID1A, we performed RNA-sequencing analysis on pre-implantation uteri from LtfiCre/+Arid1af/f and control mice. Differential expression analysis identified 4181 differentially expressed genes enriched for immune-related ingenuity canonical pathways including agranulocyte adhesion and diapedesis and natural killer cell signaling. RT-qPCR confirmed an increase in pro-inflammatory cytokine and macrophage-related gene expression but a decrease in natural killer cell signaling. Immunostaining confirmed a uterus-specific increase in macrophage infiltration. Flow cytometry delineated an increase in inflammatory macrophages and a decrease in uterine dendritic cells in LtfiCre/+Arid1af/f uteri. These findings demonstrate a role for endometrial epithelial ARID1A in suppressing inflammation and maintaining uterine immune homeostasis, which are required for successful pregnancy and gynecological health.

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OMIP‐095: 40‐Color spectral flow cytometry delineates all major leukocyte populations in murine lymphoid tissues

Kare,

Nichols,

Zermeno

et al. 2023

Cytometry Pt A

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High‐dimensional immunoprofiling is essential for studying host response to immunotherapy, infection, and disease in murine model systems. However, the difficulty of multiparameter panel design combined with a lack of existing murine tools has prevented the comprehensive study of all major leukocyte phenotypes in a single assay. Herein, we present a 40‐color flow cytometry panel for deep immunophenotyping of murine lymphoid tissues, including the spleen, blood, Peyer's patches, inguinal lymph nodes, bone marrow, and thymus. This panel uses a robust set of surface markers capable of differentiating leukocyte subsets without the use of intracellular staining, thus allowing for the use of cells in downstream functional experiments or multiomic analyses. Our panel classifies T cells, B cells, natural killer cells, innate lymphoid cells, monocytes, macrophages, dendritic cells, basophils, neutrophils, eosinophils, progenitors, and their functional subsets by using a series of co‐stimulatory, checkpoint, activation, migration, and maturation markers. This tool has a multitude of systems immunology applications ranging from serial monitoring of circulating blood signatures to complex endpoint analysis, especially in pre‐clinical settings where treatments can modulate leukocyte abundance and/or function. Ultimately, this 40‐color panel resolves a diverse array of immune cells on the axes of time, tissue, and treatment, filling the niche for a modern tool dedicated to murine immunophenotyping.

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Single-cell profiling of immune system alterations in lymphoid, barrier and solid tissues in aged mice

Cited by 21 publications

References 81 publications

Nuclear envelope disruption triggers hallmarks of aging in lung alveolar macrophages

Nuclear envelope disruption triggers hallmarks of aging in lung alveolar macrophages

Endometrial Epithelial ARID1A Is Required for Uterine Immune Homeostasis during Early Pregnancy

OMIP‐095: 40‐Color spectral flow cytometry delineates all major leukocyte populations in murine lymphoid tissues

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