Biology subjects

Ralu, M.

Publications and source records attributed to Ralu, M..

3 recordsLinked to original sources

bioRxiv2026

A Single Lipid Nanoparticle Formulation Enables Delivery of Diverse RNA Therapeutics to Human Muscle Models of Duchenne Muscular Dystrophy

Duchenne muscular dystrophy is a lethal neuromuscular disorder caused by the absence of dystrophin, for which no curative treatment is available. RNA-based approaches have shown promising results; however, their evaluation is hindered by the lack of robust and rapid delivery methods for differentiated human muscle cells, which represent the most physiologically relevant in vitro models for assessing therapeutic strategies. Here, we establish a versatile lipid nanoparticle platform enabling efficient delivery of diverse RNA therapeutics across a range of human muscle models, including myotubes, induced pluripotent stem cell-derived myotubes, myoblasts, cardiomyocytes, and 3D engineered skeletal muscle tissues. Remarkably, a single commercially available lipid nanoparticle formulation supports delivery of cargos spanning more than 300-fold in size, from short antisense oligonucleotides ([~]20 nt) to complex CRISPR-based editors (up to [~]6.7 kb), including Cas9 nucleases, adenine base editors, and CRISPRa systems. This enables efficient gene correction and transcriptional modulation, resulting in dystrophin restoration or compensatory utrophin upregulation in relevant Duchenne muscular dystrophy models. Together, our results establish a single lipid nanoparticle formulation as a versatile platform for RNA delivery in human muscle systems and provide a practical framework for the rapid preclinical assessment of emerging therapies for Duchenne muscular dystrophy and other neuromuscular disorders. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=92 SRC="FIGDIR/small/741396v1_ufig1.gif" ALT="Figure 1"> View larger version (29K): org.highwire.dtl.DTLVardef@a18d71org.highwire.dtl.DTLVardef@4d0faforg.highwire.dtl.DTLVardef@140b078org.highwire.dtl.DTLVardef@1a235_HPS_FORMAT_FIGEXP M_FIG C_FIG Created in BioRender. Leclerc, D. (2026) https://BioRender.com/90h0fme

molecular biology↗

bioRxiv2026

A combinatorial EVs-miRNA signature mediates the anti-tumoral activity of NFAT3-regulated extracellular vesicles in aggressive cancers.

Aggressive cancers such as triple-negative breast cancer (TNBC) and pancreatic cancer remain difficult to treat because their malignant behavior is driven by complex gene networks rather than single oncogenic targets. Here, we identify an extracellular vesicle (EV)-associated miRNA signature functionally linked to NFAT3 activity and demonstrate its ability to suppress tumor aggressiveness. Functional analyses revealed that a combination of fifteen miRNAs (miR-Comb 15) was required to fully reproduce the anti-tumoral effects of NFAT3-regulated EVs across TNBC and pancreatic cancer models, whereas individual miRNAs showed only partial activity. These effects were associated with coordinated regulation of validated target genes controlling proliferation and invasion, supporting a network-modulating mechanism of action. To facilitate therapeutic translation, we used EVs derived from HEK 293T cells, a non-tumoral, scalable, and readily engineerable EV source. Using an optimized exogenous pH-gradient loading strategy, miR-Comb 15 was efficiently incorporated into EVs without affecting vesicle integrity or intrinsic bioactivity. HEK 293T EVs loaded with miR-Comb 15 consistently showed the strongest anti-tumoral activity in vitro and in vivo among the delivery platforms evaluated. Together, these findings identify a functional NFAT3-dependent EV-miRNA program and support EV-mediated delivery of combinatorial miRNA therapeutics as a promising strategy for aggressive cancers.

cancer biology↗

bioRxiv2023

CRISPR-Cas9 mediated endogenous utrophin upregulation improves Duchenne Muscular Dystrophy

Duchenne muscular dystrophy (DMD) is a lethal neuromuscular disorder caused by loss of dystrophin. Upregulation of utrophin (UTRN), a dystrophin paralogue, is a promising therapeutic avenue. Here, we present a CRISPR-Cas9-mediated strategy to increase utrophin expression by disrupting microRNA (miR) binding sites (BS). Using a Cas9/gRNA ribonucleoprotein (RNP) complex we disrupted several miR BS in DMD myoblasts and selected the Let-7c BS has crucial for UTRN repression. Interestingly, Cas9/gRNA indels were as efficient as the complete removal of Let-7c BS in upregulating UTRN expression, without any major off-targets. In three-dimensional human DMD cultures, Cas9/gRNA-mediated editing resulted in significant utrophin upregulation and functional improvements of calcium dysregulation and muscle contraction. Finally, Let-7c BS disruption in mdx animals by systemic rAAVs mediated delivery of Cas9 and gRNA resulted in utrophin upregulation and amelioration of the muscle histopathological phenotype. These findings provide the foundations for a universal (mutation-independent) gene editing therapeutic strategy for DMD. One Sentence SummaryCRISPR-Cas9 has the potential to upregulate utrophin to treat all DMD patients.

molecular biology↗