Search bioRxiv⌕ Search

Biology subjects

Link, K.

Publications and source records attributed to Link, K..

3 recordsLinked to original sources

A Feature Learning Model Identifies Predictive Attributes of Mesenchymal Stromal Cell Efficacy

The therapeutic efficacy of human mesenchymal stromal cells (hMSCs) is highly variable, limiting their clinical translation for musculoskeletal diseases and other regenerative medicine applications. There is a poor understanding of the critical quality attributes correlating to therapeutic efficacy of hMSCs. To address this challenge, we analyzed pre-clinical in vitro secretome profiles and in vivo therapeutic efficacy of hMSCs from multiple human donors. hMSCs from different donors showed significant differences between donors in therapeutic efficacy when assessed in a rat post-traumatic osteoarthritis (OA) model. A partial least squares feature learning model was trained to evaluate differences between more and less therapeutic donor hMSCs by examining cytokine secretion profiles, to predict donor-specific therapeutic outcomes. More therapeutic hMSCs exhibited increased secretion of GM-CSF, GRO, IL-4, and PDGF-AA, whereas less therapeutic donors had higher TNF-, IL-6, and MCP-1 secretion. The cytokine profile was accompanied by evaluation of MAPK pathway, which revealed distinct differences in phospho-protein signaling between more and less therapeutic hMSC secretome profiles. Pharmacological inhibition of JNK signaling in more therapeutic donor cells decreased hMSC secretion of the key therapeutic associated cytokines and shifted hMSC secretome towards a less therapeutic profile. Prospective validation of cells from additional donors demonstrated significant correlations between predicted and observed pre-clinical in vivo efficacy to attenuate OA. This approach identifies critical quality attributes enabling consistent prediction of therapeutic potency, thereby addressing a major barrier to scalable and effective cell therapies. These findings advance precision cell-based therapies and offer a framework for standardized donor screening in clinical applications. SummaryA feature learning model was developed, trained, and validated to identify critical quality attributes of MSCs that predict therapeutic potency.

bioengineering↗

The non-steroidal mineralocorticoid receptor (MR) antagonist (FINERENONE) reverses Western diet-induced kidney disease by enhancing mitochondrial metabolism and decreasing lipid accumulation and inflammation

Mineralocorticoid receptor (MR) overactivation plays a crucial role in the pathogenesis of chronic kidney disease, as well as several cardiovascular and arterial diseases. Current studies determined the mechanisms of the beneficial kidney effects of the non-steroidal MR antagonist Finerenone (FN) in a mouse model of western diet-induced obesity and insulin resistance. 10-week-old male C57BL/6J mice were fed a low fat (LF) or a western diet (WD) for 12 weeks followed by treatment with either vehicle or finerenone (FN) for another 14 weeks (intervention studies) until they were 36 weeks old. Finerenone treatment prevented a) the increased albuminuria and kidney injury molecule 1 (KIM1), b) the expanded extracellular mesangial matrix, and podocyte injury, c) fibronectin, collagen IV, CD45 and CD68 immunostaining, d) glomerular basement membrane disruption, podocyte foot process loss, and mitochondrial structural abnormalities, e) the pro-inflammatory cytokines (MCP1), innate immunity pathways (TLR2, STING, STAT3), and fibrosis markers fibronectin, TGF{beta} and Pai1, and f) the increased kidney cholesterol levels. There was also reduced expression of nuclear receptor ERR{gamma} without changes in ERR in WD-fed mice whereas both ERR and ERR{gamma} expression levels increased after Finerenone treatment. NADH lifetime analysis showed decreased bound NADH, compatible with decreased mitochondrial OXPHOS in the kidneys of WD-fed mice compared to controls, which was prevented by finerenone treatment. In conclusion, Finerenone treatment exhibits a renal protective role and prevents the progression of kidney disease by regulating mitochondrial function, most likely via ERR{gamma}, and reducing lipid accumulation and inflammation.

pharmacology and toxicology↗

mRNA structure regulates protein expression through changes in functional half-life

Messenger RNAs (mRNAs) encode information in both their primary sequence and their higher order structure. The independent contributions of factors like codon usage and secondary structure to regulating protein expression are difficult to establish as they are often highly correlated in endogenous sequences. Here, we used two approaches, global inclusion of modified nucleotides and rational sequence design of exogenously delivered constructs to understand the role of mRNA secondary structure independent from codon usage. Unexpectedly, highly-expressed mRNAs contained a highly-structured coding sequence (CDS). Modified nucleotides that stabilize mRNA secondary structure enabled high expression across a wide-variety of primary sequences. Using a set of eGFP mRNAs that independently altered codon usage and CDS structure, we find that the structure of the CDS regulates protein expression through changes in functional mRNA half-life (i.e. mRNA being actively translated). This work highlights an underappreciated role of mRNA secondary structure in the regulation of mRNA stability. [150 words] HighlightsO_LIProtein expression from modified mRNAs tends to follow the pattern m1 {Psi} > U >mo5U C_LIO_LIProtein expression correlates with mRNA thermodynamic stability: {Psi}{approx} m1{Psi} > U > mo5U C_LIO_LIA highly structured CDS correlates with high expression C_LIO_LIIncreased structured mRNAs extend functional half-life C_LI

molecular biology↗