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Shaligram, S.

Publications and source records attributed to Shaligram, S..

2 recordsLinked to original sources

Ex Vivo Expanded Regulatory T Cells Inhibit AAA Progression by Limiting CD4+ and CD8+ T Cell Accumulation in Aortic Tissue

BackgroundRegulatory T cells (Tregs) play a crucial role in the pathophysiology of abdominal aortic aneurysms (AAA), a chronic inflammatory condition with few treatment options for patients with early-stage disease. Treg therapy for AAA is potentially beneficial but its specific mechanism requires further investigation for clinical applications. MethodsAfter identifying the critical role of T-cells in AAA using human and mouse AAA single-cell RNA sequencing data, we investigated the influence of Tregs on immune cell infiltration within mouse AAA--specifically CD3+ T cells--using congenic transfer of Thy1.1 allelic donor mice Tregs into AAA-induced wild-type C57BL/6J mice. AAA progression was quantified with ultrasound and image micrometry. Tissues obtained on postoperative days 7-42 were analyzed with flow cytometry, qRT-PCR, Verhoeff-van Gieson staining, hematoxylin-eosin staining, and immunohistochemistry. ResultsCD3+ T cell population was profoundly elevated in the elastase induced AAA mouse model which was further used in the study. The AAA mice that received Treg cell therapy had less elastin degradation and aortic wall enlargement than their control counterparts. Donor Tregs were detected in draining lymph nodes even after five weeks, with characteristic expression of FOXP3 and CD25. Although donor Tregs were not detected in the aortic microenvironment, the pro-inflammatory cell population including CD4 and CD8 cells was reduced, compared to control mice. ConclusionElevated T cell population aggravates inflammation and promotes AAA progression. Treg therapy impedes the recruitment of T cells into AAA tissue by colonizing the draining lymph nodes, thereby mitigating AAA progression. This study deepens our understanding of Treg stability, function, and potential as a promising therapy for early-stage aneurysms.

immunology↗

Alk1 mutant endothelial cells undergo clonal expansion in mouse brain arteriovenous malformations

RationaleMutation in human arteriovenous malformation (AVM) causative genes in a fraction of endothelial cells (ECs) causes AVMs in mice. It is unclear how a small number of mutant ECs can lead to AVM formation. ObjectiveTo understand how a fraction of mutant ECs causes AVM, we tested the following hypotheses: (1) activin receptor-like kinase 1 (Alk1 or Acvlr1) mutant brain ECs undergo clonal expansion upon angiogenic stimulation, (2) Alk1 mutant ECs display growth advantage, (3) the burden of Alk1 mutant ECs correlates with AVM severity, and (4) Alk1 mutant bone marrow (BM) derived ECs alone is sufficient to cause AVM. Methods and ResultsWe used PdgfbiCreER;Alk1f/f;confetti+/- mice which express an EC-specific tamoxifen (TM)-inducible Cre recombinase, a Cre-regulated confetti transgene, and Alk1 floxed alleles. Brain AVMs were induced by direct brain injection of an adeno-associated viral vector expressing vascular endothelial growth factor (AAV-VEGF) followed with intra-peritoneal injection of TM two weeks later. Color-predominance of confetti reporter in AVMs compared to control brain ECs suggested that clonal expansion was associated with AVM development. We treated PdgfbiCreER;Alk1f/f with different doses of TM to create a mosaic of wild-type (WT) and mutant ECs and found that equal numbers of Alk1+ and Alk1- ECs were proliferating. Increase of TM dose increased the number of Alk1- ECs, the abnormal vessels in brain AVMs, the number of arteriovenous shunts in the intestines, and mouse mortality. To test if mutation of Alk1 in BM-derived ECs can cause brain AVM, we transplanted WT mice with BM of PdgfbiCreER;Alk1f/f mice. After AAV-VEGF and TM treatment, these mice developed AVMs in their brains and arteriovenous shunts in their intestines. ConclusionClonal expansion of Alk1 mutant ECs could partly explain why a fraction of mutant ECs causes AVM. Mutation of AVM causal genes in BM-derived ECs is sufficient to cause AVM formation.

cell biology↗