Biology subjects

Lionello, V. M.

Publications and source records attributed to Lionello, V. M..

3 recordsLinked to original sources

bioRxiv2026

NAD+ limitation and α-ketoglutarate depletion define cancer induced muscle atrophy

Tumour-host crosstalk is an emerging hallmark of malignancy. It is increasingly recognised that cancer alters non-tumour bearing distant organs, with important consequences for disease progression and survival. Skeletal muscle loss, a defining feature of cancer-associated cachexia, contributes to morbidity and reduced quality of life, yet its underlying mechanisms remain poorly understood. In this study we analysed the secretome of pancreatic cancer cells and identified a metabolite-driven signal that induces myotube atrophy. Our data suggests that atrophy, which seems to involve cytoskeletal stress and increased myosin contractility, arises from cancer cell-dependent reductive metabolic rewiring. Pancreatic cancer cells conditioned their environment with elevated levels of lactate, generating a lactate-rich extracellular milieu that decrease the NAD+/NADH ratio in myotubes. This redox imbalance impaired oxidative metabolism, decreasing -ketoglutarate (KG) levels and disrupted KG-dependent processes. Restoration of NAD+ or KG levels rescued myotube atrophy. Skeletal muscle from cancer-associated cachexia mouse models exhibited reduced KG levels. Collectively, these findings identify a targetable metabolite-mediated mechanism of tumour-muscle crosstalk.

cancer biology↗

bioRxiv2025

Advanced human iPSC-based modelling of LMNA-related congenital muscular dystrophy enables development of targeted genetic therapies for muscle laminopathies

LMNA-related congenital muscular dystrophy (L-CMD) is amongst the most severe forms of laminopathies, which are diseases caused by pathogenic variants in the LMNA gene. LMNA encodes the proteins Lamin A and C, which assemble with Lamin B1 and B2 to form the nuclear lamina: a meshwork providing structural stability to the nucleus that also regulates chromatin organisation and gene expression. Research into L-CMD mechanisms and therapies is hindered by lack of humanised, tissue-specific models that accurately recapitulate disease phenotypes. We previously reported that LMNA-mutant induced pluripotent stem cell (iPSC)-derived skeletal muscle cells have nuclear shape abnormalities and Lamin A/C protein mislocalisation. Here, we expand the selection of L-CMD patient- derived iPSCs and validate disease-associated readouts using a transgene-free based protocol which more accurately mimics skeletal myogenesis. Results showed no overt defects in developmental myogenesis, but recapitulation of pathological nuclear shape abnormalities in 2D and 3D cultures, nuclear envelope protein mislocalisation and transcriptomic alterations across multiple pathogenic LMNA variants. We then utilised this platform to assess LMNA gene editing strategies. CRISPR-based exon removal generated stable RNA and protein Lamin A/C species, without significant normalisation of nuclear morphological phenotypes or transcriptomic profile. Conversely, precise editing of the same mutation showed complete reversal of disease-associated nuclear morphometrics, alongside normalisation of the pro-inflammatory transcriptomic signature. Our data provide the foundation for a humanised in vitro disease and therapy modelling platform for this complex and severe muscle disorder. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=81 SRC="FIGDIR/small/660928v2_ufig1.gif" ALT="Figure 1"> View larger version (15K): org.highwire.dtl.DTLVardef@134276forg.highwire.dtl.DTLVardef@108f44forg.highwire.dtl.DTLVardef@dfa04eorg.highwire.dtl.DTLVardef@1956a90_HPS_FORMAT_FIGEXP M_FIG C_FIG HIGHLIGHTSO_LILMNA-mutant iPSCs undergo efficient skeletal myogenesis upon transgene-free, small molecule-based lineage-directed differentiation C_LIO_LIL-CMD iPSCs recapitulate hallmark disease-associated nuclear phenotypes and show a pro-inflammatory transcriptional profile C_LIO_LIDisease modelling platforms based on iPSC-derived skeletal muscle cells enable comparative testing of gene editing strategies C_LIO_LICRISPR-edited L-CMD iPSC-derived myogenic cells show amelioration of disease-associated readouts C_LI

cell biology↗

bioRxiv2022

Combining SLA 3D printing and soft lithography for fast, versatile, and accessible high-resolution fabrication of customised multiscale cell culture devices with complex designs

Cell culture devices, such as microwells and microfluidic chips, are designed to increase the complexity of cell-based models whilst retaining control over culture conditions and have become indispensable platforms for biological systems modelling. From microtopography, microwells, plating devices and microfluidic systems to larger constructs for specific applications such as live imaging chamber slides, a wide variety of culture devices with different geometries have become indispensable in biology laboratories. However, while their application in biological projects is increasing exponentially, due to a combination of the techniques and tools required for their manufacture, and the physical science background sometimes needed, the design and fabrication of such devices directly by biological labs remains a relatively high investment in terms of costs, use of facilities, needed collaborations and time. Whilst commercially available systems are available, these are also often costly, and importantly lack the potential for customisation by each single lab. This combination of factors still limits widespread application of microfabricated custom devices in most biological wet labs. Capitalising on recent important advancements in the fields of bioengineering and microfabrication, and taking advantage of low-cost, high-resolution desktop resin 3D printers combined with PDMS soft lithography, we have developed an optimised low-cost and highly reproducible microfabrication pipeline, capable of generating a wide variety of customisable devices for cell culture and tissue engineering in an easy, fast reproducible way for a fraction of the cost of conventional microfabrication or commercial alternatives. This protocol is designed specifically to be a resource for biological labs with little to none prior exposure to these fields technique and enables the manufacture of complex devices across the {micro}m to cm scale. We provide a ready-to-go pipeline for the efficient treatment of resin-based 3D printed constructs for PDMS curing, using a combination of curing steps, washes and surface treatments. Together with the extensive characterisation of the fabrication pipeline, we show the utilization of this system to a variety of applications and use cases relevant to biological experiments, ranging from micro topographies for cell alignments to complex multi-part hydrogel culturing systems. This methodology can be easily adopted by any wet lab, irrespective of prior expertise or resource availability and will enable the wide adoption of tailored microfabricated devices across many fields of biology.

bioengineering↗