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2. It’s what’s on the inside that counts: capsid content as a critical quality attribute for AAV characterizationLauriel F. Earley, Stuart Nelson, Ana Dolinar Češarek, David Dobnik, 2026, other scientific articles Abstract: Recombinant adeno-associated virus (rAAV) vectors exhibit complex heterogeneity in capsid content. Although a growing array of analytical platforms can interrogate different capsid attributes, no single method provides a comprehensive or unequivocal assessment. Critically, current approaches often fail to evaluate genome integrity, an emerging important quality attribute. Incomplete genomes can contribute to viral genome titers without conferring therapeutic benefit, while simultaneously increasing immunogenic burden and risk of adverse events. Discordant results from orthogonal assays further complicate interpretation and cross-laboratory comparisons. Here, we highlight the limitations of existing characterization strategies and advocate for robust analytical frameworks that directly assess genome completeness. We propose best-practice guidelines, including improved genome integrity assays and harmonized reporting, to better link capsid content analytics with potency and patient safety in gene therapy development.
Keywords: gene therapy, AAV, genomic integrity, capsid content
Published in DiRROS: 23.06.2026; Views: 262; Downloads: 189
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3. The genetic blueprint of obesity : from pathogenesis to novel therapiesGašper Tonin, Stjepan Eržen, Zala Mlinarič, Dubravka Jurišić-Eržen, Simon Horvat, Tanja Kunej, Jasna Klen, 2025, review article Keywords: CRISPR/Cas9, dual therapy, energy homeostasis, gene therapy, gut–brain axis, microbiota, personalized medicine
Published in DiRROS: 22.04.2026; Views: 278; Downloads: 284
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4. Eculizumab for thrombotic microangiopathy induced by onasemnogene abeparvovec in spinal muscular atrophyTanja Kersnik-Levart, Nina Olas Kar, Chiara Močnik Pegan, Eva Vrščaj, Anja Troha Gergeli, Tanja Loboda, Damjan Osredkar, 2025, other scientific articles Abstract: Introduction: Onasemnogene abeparvovec is one of the three disease-modifying therapies available that can significantly improve the outcome of patients with 5q-spinal muscular atrophy. Therapy-induced thrombotic microangiopathy is an ultra-rare, but potentially life-threatening condition of not yet clearly defined aetiology. Case Presentation: A case of a 2-year-old patient with 5q-spinal muscular atrophy, who developed thrombotic microangiopathy after gene replacement therapy with onasemnogene abeparvovec, is described. This severe adverse event was promptly recognized and successfully treated with the complement C5 inhibitor. Conclusion: Thrombotic microangiopathy is an ultra-rare, but potentially life-threatening condition that can occur after onasemnogene abeparvovec therapy. Anticipation of these serious adverse events, its prompt recognition and treatment is crucial for a better outcome.
Keywords: spinal muscular atrophy, gene replacement therapy, onasemnogene abeparvovec, thrombotic microangiopathy, eculizumab
Published in DiRROS: 10.03.2026; Views: 574; Downloads: 353
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5. Chemokine CCL5 overexpression combined with radiotherapy modulates Th1-mediated immune response and leads to significant tumor growth delay in mouse tumor modelsTim Božič, Iva Šantek, Živa Pišljar, Simona Kranjc Brezar, Gregor Serša, Boštjan Markelc, Maja Čemažar, 2026, original scientific article Abstract: This study investigated the antitumor efficacy of chemokine CCL5 gene therapy using gene electrotransfer (GET) in combination with radiotherapy (RT) in solid murine tumors CT26 and 4T1. In vitro, CT26 and 4T1 tumor cells transfected with plasmid DNA (pDNA) encoding CCL5 induced migration of RAW264.7 macrophages. In vivo, CCL5 overexpression achieved via GET of pDNA encoding CCL5 led to increased splenocyte infiltration in dorsal window chamber models. When combined with RT, GET of pDNA encoding CCL5 shifted the tumor cytokine profile toward a proinflammatory state, with elevated Ifn-γ, Cxcl9, Cxcl10, and Il-12α. Although CD8 + and CD4 + T cells were reduced post-treatment, due to radiation-induced cell death, the combination of GET of pDNA encoding CCL5 and RT significantly delayed tumor growth in both models. In 4T1 tumors, this delay was also significant compared to the equivalent treatment with GET of control pDNA. These findings support GET of pDNA encoding CCL5 combined with RT as a strategy to enhance immune-mediated tumor control.
Keywords: chemokines, gene electrotransfer, gene therapy, mouse, radiotherapy
Published in DiRROS: 13.02.2026; Views: 654; Downloads: 226
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6. Between defence and delivery : the DNA sensing response to gene electrotransferTanja Jesenko, Maša Omerzel, Loree C. Heller, Maja Čemažar, 2025, review article Abstract: Gene therapy has emerged as a transformative biomedical approach, offering new therapeutic possibilities from many so far uncurable diseases through the introduction of recombinant nucleic acids into target cells. Among non-viral delivery techniques, gene electrotransfer (GET) has become one of the frequently applied methods in clinical trials. It is based on the application of short, high-intensity electric pulses that transiently permeabilize cell membranes and enable the efficient transfer of plasmid DNA or other types of recombinant nucleic acids into various cell types. Beyond its role in gene delivery, GET can trigger complex cellular responses, as the introduced DNA interacts with intracellular DNA sensing pathways involved in innate immunity and inflammation. These responses can influence the therapeutic outcome – either by enhancing antitumour and vaccine-related immune activation or by reducing transfection efficiency when excessive inflammation or cell death occur. Our experimental findings in tumour, muscle, and skin models have shown that even non-coding plasmid DNA delivered by GET can induce local immune stimulation and tissue-specific inflammatory signaling, suggesting that the delivered DNA itself contributes to therapeutic efficacy. Conclusions The dual nature of cellular responses following plasmid DNA GET represents both an opportunity and a challenge. Controlled activation of innate immunity can be harnessed to amplify antitumour or vaccine efficacy, while excessive responses may hinder applications requiring cell survival and sustained expression. Understanding these mechanisms enables the rational optimization of GET parameters and plasmid vector design to fully exploit the adjuvant effect or reduce the off-target effect of DNA sensing after GET, based on the desired application.
Keywords: gene electrotransfer, DNA sensors, gene therapy
Published in DiRROS: 16.01.2026; Views: 770; Downloads: 279
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7. Gene therapy of rare diseases as a milestone in medicine : overview of the field and report on initial experiences in SloveniaUrh Grošelj, Marko Kavčič, Ana Drole Torkar, Jan Kafol, Duško Lainšček, Roman Jerala, Matjaž Sever, Samo Zver, Gregor Serša, Maja Čemažar, Primož Strojan, Aleš Grošelj, Mojca Žerjav-Tanšek, Špela Miroševič, Simona Ivančan, Tomaž Prelog, David Gosar, Jasna Oražem, Matej Mlinarič, Sara Bertok, Jernej Kovač, Jana Kodrič, Saba Battelino, Marko Pokorn, Alojz Ihan, Janez Jazbec, Tadej Battelino, Damjan Osredkar, 2025, review article Abstract: Gene therapy has transitioned from a long-awaited promise to a clinical reality, offering transformative treatments for rare congenital diseases and certain cancers, which have a significant impact on patients’ lives. Current approaches focus on gene replacement therapy, either in vivo or ex vivo, mostly utilizing viral vectors to deliver therapeutic genes into target cells. However, refining these techniques is essential to overcome challenges and complications associated with gene therapy to ensure long-term safety and efficacy. Slovenia has witnessed significant advancements in this field since 2018, marked by successful gene therapy trials and treatments for various rare diseases. Significant strides have been made in the field of gene therapy in Slovenia, treating patients with spinal muscular atrophy and rare metabolic disorders, including the pioneering work on CTNNB1 syndrome. Additionally, immune gene therapy, exemplified by IL-12 adjuvant therapy for cancer, has been a focus of research in Slovenia. Through patient-centred initiatives and international collaborations, researchers in Slovenia are advancing preclinical research and clinical trials, paving the way for accessible gene therapies. Establishing clinical infrastructure and genomic diagnostics for rare diseases is crucial for gene therapy implementation. Efforts in this regard in Slovenia, including the establishment of a Centre for Rare Diseases, Centre for the Technologies of Gene and Cell Therapy, and rapid genomic diagnostics, demonstrate a commitment to comprehensive patient care. Despite the promises of gene therapy, challenges remain, including cost, distribution, efficacy, and long-term safety. Collaborative efforts are essential to address these challenges and ensure equitable access to innovative therapies for patients with rare diseases.
Keywords: gene therapy, rare genetic diseases, Slovenia, CAR-T cells, cancer, immune gene therapy
Published in DiRROS: 04.12.2025; Views: 746; Downloads: 459
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8. Unravelling genetic etiology of cerebral palsy : findings from a Slovenian pediatric cohortUla Arkar Silan, Ana Trebše, Jernej Kovač, Mihael Rogač, Anja Troha Gergeli, Robert Šket, Tina Bregant, David Neubauer, Borut Peterlin, Damjan Osredkar, 2025, original scientific article Abstract: Introduction: Cerebral palsy (CP) is a permanent movement or postural disorder due to non-progressive injury to the developing brain, with recent research suggesting a genetic contribution in many patients. This study aimed to investigate the genetic etiology of CP in Slovene children without a previously suspected genetic cause or with prior negative genetic testing. Methods: All children born after 2003 from the Slovenian National Registry of Cerebral Palsy (SRCP) without an established genetic diagnosis were invited to participate in this cross-sectional study. Whole exome sequencing (WES) was conducted, followed by analysis of 110 CP-associated genes. Thirteen patients underwent additional family segregation by Sanger sequencing. Genetic findings were classified according to the ACMG guidelines. Results: The study included 136 children, of whom 68 (50%) were male. Spastic CP was identified in 85% of the participants, dyskinetic in 13%, and ataxic in 2%. Gross Motor Function Classification System (GMFCS) levels varied, with the majority (36%) classified as level I. Pathogenic variants, likely pathogenic variants, or ‘de novo’ variants of unknown significance (VUS) were identified in nine children (6.6%) in ATL1, CTNNB1, DYRK1, KMT2A, PROC, SPAST, ZC4H2, and ZSWIM6. Among these nine children, two had normal brain Magnetic Resonance Imaging (MRI) and three had an unsuspicious medical history. Conclusion: This study identified plausible, possible, or definite genetic etiologies in a cohort of children with CP. Apart from the exclusion of individuals with a previously established genetic diagnosis, no other selection criteria were applied, allowing for an inclusive assessment of genetic contributions within this population. With the advent of personalized medicine and genetic treatment, understanding the genetic underpinnings of CP is crucial for targeted therapy.
Keywords: cerebral palsy, genetic etiology, whole exome sequencing, gene therapy, CTNNB1
Published in DiRROS: 21.11.2025; Views: 665; Downloads: 325
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