表达抗纤维化载物的基因工程人骨髓间充质基质细胞的肾保护疗效和安全性。

The renoprotective efficacy and safety of genetically-engineered human bone marrow-derived mesenchymal stromal cells expressing anti-fibrotic cargo.

机构信息

Cardiovascular Disease Program, Monash Biomedicine Discovery Institute, Monash University, Clayton, VIC, 3800, Australia.

Department of Pharmacology, Monash University, Clayton, VIC, 3800, Australia.

出版信息

Stem Cell Res Ther. 2024 Oct 23;15(1):375. doi: 10.1186/s13287-024-03992-x.

DOI:10.1186/s13287-024-03992-x

PMID:39443975

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11515549/

Abstract

BACKGROUND

Kidney fibrosis is a hallmark of chronic kidney disease (CKD) and compromises the viability of transplanted human bone marrow-derived mesenchymal stromal cells (BM-MSCs). Hence, BM-MSCs were genetically-engineered to express the anti-fibrotic and renoprotective hormone, human relaxin-2 (RLX) and green fluorescent protein (BM-MSCs-eRLX + GFP), which enabled BM-MSCs-eRLX + GFP delivery via a single intravenous injection.

METHODS

BM-MSCs were lentiviral-transduced with human relaxin-2 cDNA and GFP, under a eukaryotic translation elongation factor-1α promoter (BM-MSCs-eRLX + GFP) or GFP alone (BM-MSCs-eGFP). The ability of BM-MSCs-eRLX + GFP to differentiate, proliferate, migrate, produce RLX and cytokines was evaluated in vitro, whilst BM-MSC-eRLX + GFP vs BM-MSCs-eGFP homing to the injured kidney and renoprotective effects were evaluated in preclinical models of ischemia reperfusion injury (IRI) and high salt (HS)-induced hypertensive CKD in vivo. The long-term safety of BM-MSCs-RLX + GFP was also determined 9-months after treatment cessation in vivo.

RESULTS

When cultured for 3- or 7-days in vitro, 1 × 10 BM-MSCs-eRLX + GFP produced therapeutic RLX levels, and secreted an enhanced but finely-tuned cytokine profile without compromising their proliferation or differentiation capacity compared to naïve BM-MSCs. BM-MSCs-eRLX + GFP were identified in the kidney 2-weeks post-administration and retained the therapeutic effects of RLX in vivo. 1-2 × 10 BM-MSCs-eRLX + GFP attenuated the IRI- or therapeutically abrogated the HS-induced tubular epithelial damage and interstitial fibrosis, and significantly reduced the HS-induced hypertension, glomerulosclerosis and proteinuria. This was to an equivalent extent as RLX and BM-MSCs administered separately but to a broader extent than BM-MSCs-eGFP or the angiotensin-converting enzyme inhibitor, perindopril. Additionally, these renoprotective effects of BM-MSCs-eRLX + GFP were maintained in the presence of perindopril co-treatment, highlighting their suitability as adjunct therapies to ACE inhibition. Importantly, no major long-term adverse effects of BM-MSCs-eRLX + GFP were observed.

CONCLUSIONS

BM-MSCs-eRLX + GFP produced greater renoprotective and therapeutic efficacy over that of BM-MSCs-eGFP or ACE inhibition, and may represent a novel and safe treatment option for acute kidney injury and hypertensive CKD.

摘要

背景

肾纤维化是慢性肾脏病（CKD）的标志，并损害了移植的人骨髓间充质基质细胞（BM-MSCs）的活力。因此，BM-MSCs 被遗传工程改造以表达抗纤维化和肾保护激素人松弛素-2（RLX）和绿色荧光蛋白（BM-MSCs-eRLX+GFP），这使得 BM-MSCs-eRLX+GFP 能够通过单次静脉注射进行传递。

方法

BM-MSCs 通过慢病毒转导带有人类松弛素-2 cDNA 和 GFP，在真核翻译延伸因子-1α启动子（BM-MSCs-eRLX+GFP）或 GFP 单独（BM-MSCs-eGFP）下。评估了 BM-MSCs-eRLX+GFP 在体外分化、增殖、迁移、产生 RLX 和细胞因子的能力，而在缺血再灌注损伤（IRI）和高盐（HS）诱导的高血压 CKD 的临床前模型中评估了 BM-MSCs-eRLX+GFP 归巢至损伤肾脏和肾保护作用。还在体内治疗停止后 9 个月确定了 BM-MSCs-RLX+GFP 的长期安全性。

结果

当在体外培养 3 或 7 天时，1×10 BM-MSCs-eRLX+GFP 产生了治疗性 RLX 水平，并分泌了增强但精细调节的细胞因子谱，而不会损害其增殖或分化能力与原始 BM-MSCs 相比。BM-MSCs-eRLX+GFP 在给药后 2 周被鉴定在肾脏中，并在体内保留了 RLX 的治疗作用。1-2×10 BM-MSCs-eRLX+GFP 减轻了 IRI 或治疗性阻断的 HS 诱导的肾小管上皮损伤和间质纤维化，并显著降低了 HS 诱导的高血压、肾小球硬化和蛋白尿。与单独给予 RLX 或 BM-MSCs 相比，其效果相当，但比 BM-MSCs-eGFP 或血管紧张素转换酶抑制剂培哚普利的效果更广泛。此外，在培哚普利联合治疗的情况下，BM-MSCs-eRLX+GFP 的这些肾保护作用得以维持，这突出了它们作为 ACE 抑制的辅助治疗的适用性。重要的是，没有观察到 BM-MSCs-eRLX+GFP 的主要长期不良影响。