Citations

Abstract Importance Mesenchymal stromal cells (MSCs) possess therapeutic properties that mediate repair. Obesity impairs MSC functionality and therapeutic efficacy, possibly by eliciting dynamic modifications of epigenetic markers, like 5-hydroxymethylcytosine (5hmC). Objective We hypothesized that human obesity alters the 5hmC landscape and anti-inflammatory capacity of adipose tissue-derived MSCs to activate the prominent inflammatory signaling mediator Interleukin (IL)-1β. Design, Setting, Participants, Intervention Adipose tissue samples were collected from obese and lean individuals (body mass index ≥30 or <30 kg/m2, respectively, n = 11 each) during weight-loss or kidney donation surgery. Main Outcomes and Measures MSCs were harvested and analyzed for 5hmC profiles (MeDIP-seq) and mRNA expression (RNA-seq) (n = 5 each). Subsequently, MSCs or a vehicle were injected into mice, (n = 6 each) and two-weeks later, kidneys were evaluated using in-vivo magnetic resonance imaging and ex vivo studies. The role of IL-1β was then studied in-vitro in MSC-induced immunomodulation using siRNA in macrophages. Results Compared to MSC from lean patients, obese-MSC genes showed 2087 differential 5hmC modifications and 175 differential mRNA expression. Among them, 14 genes with overlapping alterations were involved in regulation of cytokine production, prominently IL-1β. Injecting obese MSCs elevated renal expression of IL-1β and M1 macrophage count but lowered kidney perfusion. Silencing IL-1β in obese-MSCs in vitro reduced M1 phenotype switching in co-incubated macrophages. Conclusions and Relevance Obesity induces epigenetic and gene expression changes in MSCs, particularly in IL-1β, associated with impaired anti-inflammatory functionality of MSCs. Targeting IL-1β could be a useful therapeutic approach to modulate the decline in MSC functionality resulting from obesity.

Key Points We characterized the renal transcriptional profile at the single-nuclear level in a preclinical swine model of CKD. Our study reveals that renal endothelial cells represent the cluster with the highest number of differentially expressed genes related to inflammatory and fibrotic activity. These results offer new insights into CKD-induced transcriptional changes in renal endothelial cells at the single-nuclear level. Background The prevalence of CKD is on the rise, and precision strategies to offset the progression to ESKD are needed. We used a well-established preclinical translational model of CKD in swine and single-nucleus RNA sequencing (snRNA-seq) to identify potential therapeutic targets through characterization of the renal cell-specific transcriptomic landscape. Methods Normal and CKD pigs were studied in vivo and ex vivo after 14 weeks ( n =6/group). In randomly selected pigs ( n =3/group), kidneys were harvested, nuclei isolated, libraries prepared, and snRNA-seq performed. Protein expression of candidate differentially expressed genes (DEGs, log2FC >0.25, adjusted P value < 0.05) was determined by immunohistochemistry, and their expression in primary normal and CKD renal vascular endothelial cells was modulated (small interfering RNA) in vitro . Results A total of 52,213 nuclei were analyzed. Thirty clusters were identified and filtered by canonical gene markers, revealing 16 unique renal cell types. Endothelial cells were the top cell type exhibiting the highest number of DEGs, which were subsequently filtered by angiogenesis, inflammation, and fibrosis (major injurious pathways altered in CKD kidneys). Venn diagram analysis identified five unique overlapping DEGs in endothelial cells: vWF , LAMA3 , and KDR upregulated and PTGIS and ICAM1 downregulated in CKD versus normal kidneys. Venn diagram analysis indicates that vWF , LAMA3 , and ICAM1 participate in inflammatory and fibrotic signaling. Renal protein expression of these DEGs matched snRNA-seq findings. Furthermore, in vitro silencing of vWF and LAMA3 ameliorated endothelial cell inflammatory and fibrotic signaling. Conclusions Our work characterized the single-nuclear renal cell transcriptomic landscape of a translational model of CKD and singled out genes implicated in major renal injury pathways. These genes could serve as potential targets to pave the way for new therapeutic strategies in patients with CKD.

Objectives: To analyze variations in patient characteristics, treatment patterns, clinical manifestations, clinical outcomes (i.e., response, flares and flare-free survival in HES; remission, relapses and relapse-free survival in EGPA; and overall survival), and healthcare resource utilization (HCRU) in patients with hypereosinophilic syndrome (HES) and eosinophilic granulomatosis with polyangiitis (EGPA) across five European countries. Methods: In two parallel, retrospective, non-interventional, longitudinal chart review studies (GSK ID: 214661 [EGPA] and 214657 [HES]), physicians collected data of patients they treated in France, Germany, Italy, Spain, and the UK from January 2015 to December 2019, with follow-up until August 2021. Country-level results are presented for each study; HES and EGPA data were pooled in a post hoc exploratory analysis. Results: Per-country, 22–26 HES- and 38–45 EGPA-treating physicians collected data from 52–62 (total 280) patients with HES and 80–85 (total 407) with EGPA. Patient sex and age at diagnosis differed across countries. Pooled HES/EGPA data revealed high oral corticosteroid (OCS) use in all countries (94.9% of patients; median [IQR] duration 20.7 [9.0, 33.8] months); immunosuppressive treatments and biologics use varied between countries (43.7–61.5% and 25.6–59.8%, respectively). The most frequent clinical manifestations were constitutional (51.6–78.8%) and lung (43.5–55.8%) in HES, and lung (41.3–67.9) and ENT (43.8–61.2) in EGPA. Pooled HCRU data showed country-level variation; 70.7–91.2% of patients had disease-related outpatient visits and 36.1–52.6% had ER visits or hospitalizations. Conclusions: Results demonstrate substantial disease burden, including high HCRU and extensive OCS use among patients with HES and EGPA in five European countries. The findings highlight the need for improved treatment strategies such as optimizing use of biologics to mitigate the reliance on corticosteroids.

Abstract Renal artery stenosis (RAS) is the leading cause of secondary hypertension worldwide. However, current medical and surgical treatment modalities provide minimal benefits for kidney injury. Recent preclinical RAS models have demonstrated promising potential of human mesenchymal stem cells (MSC) and their daughter extracellular vesicles (EV) in improving murine renal function and attenuating inflammation. However, the extent and mechanisms underlying immune rejection of xenogeneic MSCs or EVs are yet undetermined. Therefore, adipose tissue was harvested from adult healthy patients. Adipose-derived MSCs were extracted and cultured, and EVs were isolated from their supernatants via ultra-centrifugation. Then, mice randomly assigned to RAS or sham surgery were divided into 6 groups: sham surgery, RAS, sham + MSC, RAS + MSC, sham + EV, and RAS + EV. Two weeks after intra-aortic injection of MSCs (5 × 105) or EVs (20 µg protein), we compared the intrarenal T-cell and macrophage accumulation, splenic B-cell numbers, circulating cytokines and anti-human antibodies levels among the groups. MSCs and EVs did not influence intrarenal immune cell infiltrations. However, MSCs significantly increased circulating anti-human antibodies. In the spleen, RAS + EV mice showed higher memory IgM+ B-cells but reduced CD19+ B-cells compared to RAS + MSC. In vitro T-cell recall assay showed that both MSCs and EVs exhibited reduced IFN-γ release upon re-stimulation, indicating an immunosuppressive effect. Therefore, xenogeneic MSCs induced a greater humoral response in mice, while EVs triggered a splenic cellular response, but neither elicits discernible kidney rejection. Our results provide key insights into the immunomodulatory mechanisms of MSCs and EVs and immune mechanisms underlying xenograft rejection.

ABSTRACTBackground and PurposeLarge‐bore aspiration catheters (LBACs) are used for thrombectomy in large vessel occlusion (LVO), either as a standalone direct aspiration first‐pass technique or combined with a stent retriever (ASR). LBAC design may influence ASR thrombectomy efficacy. We compared the safety and performance of the novel MIVI Q segmental catheter with the well‐established SOFIA aspiration device in ASR thrombectomy.MethodsWe analyzed data from the Registry cOmbined vS SinglE Thrombectomy TechnIques registry of consecutive patients with anterior circulation LVO and compared the outcomes of those treated with first‐line ASR thrombectomy using Q (Q5 or Q6) or SOFIA (5F or 6F Plus) catheters. Demographic, clinical, angiographic, and clinical outcome data (24‐h National Institute of Health Stroke Scale [NIHSS] and modified Rankin Scale score at 3 months) were compared.ResultsOf the 853 patients, 155 (18.2%) were treated with MIVI Q and 698 (81.8%) with SOFIA catheters. After adjusting for age, sex, NIHSS score at baseline, tPA use, site occlusion, anesthesia type, and diameter and length of SR, the MIVI Q group was comparable to the SOFIA group in terms of first‐pass effect or successful final recanalization and safety. However, the MIVI Q group had a shorter mechanical thrombectomy time (20 [10–45] min vs. 33 [20–51] min; odds ratio [OR] = 7.4, 95% confidence interval [CI]: 1.1–14; p = 0.021) and a lower rate of symptomatic intracerebral hemorrhage (3.3% vs. 8.8%; OR = 3.59, 95% CI: 1.45–10.9; p = 0.011).ConclusionsIn ASR neurothrombectomy, SOFIA aspiration catheters were not superior to MIVI Q in achieving successful and complete first‐passage recanalization; however, MIVI Q had shorter procedural times and a lower rate of symptomatic intracranial hemorrhage.