Functional validation of allele-specific LMNB1 silencing in patient-derived astrocytes as a therapeutic option for Autosomal Dominant Leukodystrophy
Year: 2026
Type: preprint
Abstract: Adult-onset Autosomal Dominant Leukodystrophy (ADLD) is a rare fatal leukodystrophy caused by increased LMNB1 gene dosage, most commonly resulting from duplication of the LMNB1 locus. Because ADLD is a gene dosage disorder, selective reduction of pathological LMNB1 expression represents a rational therapeutic strategy. Although allele-specific RNA interference has previously been shown to lower LMNB1 levels in patient-derived fibroblasts and directly reprogrammed neurons, its therapeutic effects have not been evaluated in disease-relevant human glial cells or using functional efficacy endpoints. Here, we established human induced pluripotent stem cell-derived astrocytes from ADLD patients as a human glial model in which to validate allele-specific LMNB1 silencing across molecular, cellular, and functional readouts. ADLD astrocytes recapitulated increased LMNB1 expression and characteristic nuclear abnormalities and displayed transcriptional alterations affecting extracellular matrix organization, calcium homeostasis, metabolism and RNA processing. Functionally, these cells also exhibited functional phenotypes suitable for therapeutic evaluation: astrocyte-conditioned medium impaired the viability of both murine and human oligodendroglial cultures, while conditioned-medium and direct astrocyte-seeding paradigms revealed impaired post-lesion myelin recovery in lysolecithin-treated cerebellar organotypic slices. Allele-specific LMNB1 silencing restored physiological LMNB1 levels, corrected nuclear abnormalities, attenuated astrocyte-mediated oligodendroglial toxicity, improved post-lesion myelin recovery, and was associated with selective transcriptional programs associated with extracellular support and cholesterol metabolism. Together, these findings provide molecular, cellular, and functional validation of allele-specific LMNB1 dosage correction in patient-derived human astrocytes and offer key support for LMNB1-lowering strategies in disease-relevant human glial cells.
Authors: Martina Lorenzati, Arianna Contato, Fernando Josa-Prado, Marta Ribodino, Maryam K. Ardakani, Ersilia Nicorvo, Elena Signorino, Giacomo Turrini, Valentina Cerrato, Roberta Parolisi, Filippo Lucchini, Gabriele Piacenti, Giacomo Donati, Valentina Proserpio, Juan Antonio Garcìa-Leòn, Marzia Rossato, Luciano Conti, Alfredo Brusco, Giulia Ramazzotti, Stefano Ratti, Pietro Cortelli, Elisa Giorgio, Annalisa Buffo
Institutions: Neuroscience Institute Cavalieri Ottolenghi, University of Turin, University of Verona, Biomedical Research Networking Center on Neurodegenerative Diseases, Instituto de Investigación Biomédica de Málaga, Universidad de Málaga, University of Trento, University of Bologna, University of Pavia
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: green
Awards: G3323308467, G8142160590
OpenAlex ID: W7213320339