European Journal of Translational Myology
The European Journal of Translational Myology (previously published as Basic and Applied Myology) is recognized as the authoritative European forum in the field of Translational Myology. The journal provides a comprehensive platform for the dissemination of high-quality research addressing muscle biology and pathology across multiple levels of investigation, including fundamental, cellular, molecular, genetic, clinical, and translational studies. Its scope encompasses a broad array of article types, such as original research papers, systematic and narrative reviews, editorials, viewpoints, methodological protocols, rapid communications, comments, and letters to the editor. Through this multifaceted approach, the journal fosters scholarly exchange and advances the integration of basic and applied knowledge into clinical and translational practice.
PAGEPress Scientific Publications, Pavia, Italy
PAGEPress has chosen to apply the Creative Commons Attribution NonCommercial 4.0 International License (CC BY-NC 4.0) to all manuscripts to be published.
Anterolateral approach for total ankle arthroplasty, preliminary results of a less-invasive surgical pathway: an observational retrospective study
Total Ankle Arthroplasty (TAA) has evolved into a reliable treatment option for end-stage ankle osteoarthritis; however, wound healing complications remain a significant concern, particularly with traditional anterior approaches. These complications are largely related to the vulnerability of the anterior soft-tissue envelope and may negatively affect early outcomes. The purpose of the present study was to evaluate the safety and early clinical outcomes of a curvilinear anterolateral approach for TAA, with particular focus on wound healing and soft-tissue complications. A retrospective observational study was conducted on 48 consecutive patients undergoing TAA between January 2023 and December 2025. A total of 49 ankle arthroplasties were performed using a curvilinear anterolateral approach. Patient demographics, comorbidities, and perioperative variables were recorded. Primary outcomes included wound-related complications and time to complete healing. Secondary outcomes included functional improvement assessed using the Foot and Ankle Outcome Score (FAOS). Three patients (6.1%) developed wound-related complications. Two patients (4.1%) experienced superficial wound dehiscence, which resolved with conservative treatment. One patient (2.0%) developed a deeper wound complication involving the soft tissues without prosthetic involvement; this case occurred in a high-risk patient with multiple comorbidities and was successfully managed with advanced wound care. No cases of deep prosthetic infection were observed. All remaining cases (93.9%) achieved uneventful wound healing within 12 to 21 days. Functional outcomes showed a significant improvement, with mean FAOS increasing from 40 ± 5 preoperatively to 75 ± 5 at final follow-up. The curvilinear anterolateral approach for TAA demonstrated a low rate of wound-related complications and consistent early functional improvement. By preserving the tibialis anterior tendon sheath and optimizing incision placement, this technique may reduce both vascular and mechanical risk factors associated with wound healing disorders. It represents a safe and reproducible alternative to traditional anterior and transfibular approaches, particularly in patients at increased risk for soft-tissue complications.
Copyright (c) 2026 the Author(s) http://creativecommons.org/licenses/by-nc/4.0
Does physical therapist contact matter? A randomised controlled trial comparing telerehabilitation and booklet-based exercise for knee osteoarthritis
Telerehabilitation is increasingly used in knee osteoarthritis, but its effectiveness compared with structured home exercise and the role of physical therapist contact remain unclear. To address this, we aimed to investigate whether the clinical effectiveness of telerehabilitation for knee osteoarthritis is primarily attributable to the digital platform itself or to the extent and quality of physical therapist engagement within that platform. In this single-centre Randomised Controlled Trial (RCT), 118 patients with knee OA (Kellgren–Lawrence grade I–III) were allocated 1:1 to a 12-week telerehabilitation (TR; n=57) or booklet-based exercise programme (BB; n=61). Within the telerehabilitation group, participants were retrospectively stratified into those with Physical Therapist Contact (TR-C) and those without contact (TR-NC) for an exploratory analysis. Outcomes included knee pain (NRS, Numerical Rating Scale), Range Of Motion (ROM), WOMAC, physical activity (IPAQ), and health-related quality of life (SF-12). A post-hoc exploratory subgroup analysis was conducted and looked at the impact of a physical therapist with and without contact. A key component of the analysis was an exploratory subgroup comparison examining the role of physical therapist contact within the telerehabilitation group. Primary outcomes were analysed using mixed-model ANOVA, with exploratory subgroup comparisons. Both groups showed significant within-group improvements in pain, ROM, and WOMAC scores (all p ≤0.001), with no significant group × time interactions (p >0.05). Exploratory analysis identified significant differences based on physical therapist contact, with greater improvements in pain (p=0.009), knee flexion (p=0.041), and WOMAC Pain (p=0.047) among participants who engaged with the physical therapist. Participants without contact showed the smallest improvements. When stratified by physical therapist contact, participants receiving contact demonstrated greater improvements than those without contact, who showed the smallest gains. Telerehabilitation and booklet-based exercise programme provide comparable clinical outcomes at the group level. However, physical therapist engagement appears to be a key determinant of treatment response. These findings suggest that the effectiveness of TR depends more on clinical interaction than on digital delivery alone. Physical therapist contact may represent an important component influencing the effectiveness of telerehabilitation. It remains unclear whether telerehabilitation effectiveness stems from the digital platform, exercise content, or physical therapist guidance.
Many excellent and one bad news regarding the European Journal of Translational Myology and the 2027 Padua Days on Muscle and Mobility Medicine
Recently, both Clarivate and Scopus announced improved metrics for the European Journal of Translational Myology (EJTM), bringing its Impact Factor to 2.0 and its CiteScore to 4.1. I am proud of these achievements and share the credit with the entire PAGEpress staff. Furthermore, many organizers and moderators for the "Padua Days on Muscle and Mobility Medicine" 2027 (2027 Pd3m) have accepted their roles and proposed improvements for the event, which will take place at the Euganean Thermal Baths (Padua, Italy) from March 9 to 12, 2027. The program remains open to numerous additional speakers. On the downside, rising operating costs are affecting both digital publishing and event organization; it is hoped that these increases will not negatively impact the submission of contributions to the EJTM and the 2027 Pd3m conference. Those interested can find further information and forms for active participation (registration, accommodation, abstract template, and participation details) at the following address: www.paduamuscledays.it. Undoubtedly, the 2027 Pd3m promises to be an engaging event, on par with all previous editions of the "Padua Days on Muscle and Mobility Medicine."
Clinical efficacy of ramp lesion treatment: a systematic review and meta analysis study
Ramp lesions, tears involving the peripheral attachment of the posterior horn of the medial meniscus, significantly impact knee stability and function, particularly in association with anterior cruciate ligament injuries. However, evidence regarding optimal treatment strategies remains inconclusive. This systematic review and meta-analysis aimed to evaluate and compare the clinical efficacy of various treatment modalities for ramp lesions. A comprehensive systematic search was conducted across major electronic databases, including PubMed, Scopus, and Web of Science, up to June 2025. Studies were included based on predefined eligibility criteria. Data extraction was performed and meta-analytic synthesis was conducted using random-effects models. A total of 17 studies met inclusion criteria. The pooled mean difference demonstrated significant improvements in both IKDC (MD=28.5 points; 95% CI: 27.9–29.2) and Lysholm scores (MD=28.5 points; 95% CI: 27.9–29.2). Subgroup analyses revealed notable effectiveness for AIR compared to CM and PM, with statistically significant subgroup differences (p<0.05). The overall treatment success rate was 87% (95% CI: 83–92%). Across techniques, patient-reported outcomes improved markedly (IKDC fixed-effect pooled Mean Difference [MD] 28.8 points, 95% CI 28.2–29.4; random-effects overall effect significant; I²=98%). In anterior vs posterior all-inside subgroups, IKDC showed no robust between-approach difference on the random-effects test (χ²=0.51, p=.48), with a small fixed-effect difference (χ²=4.34, p=.04). Lysholm scores also improved substantially (fixed-effect pooled MD 28.0 points, 95% CI 27.3–28.7), and the between-approach difference was significant, favoring the posterior subgroup on the fixed-effect model. The pooled clinical success proportion was 90% without a significant difference between approaches (p=.10). The findings indicate that surgical interventions, particularly All-inside repair has effectively improved knee function and patient-reported outcomes following ramp lesions compared to conservative management and partial meniscectomy. However, no
Acute beetroot juice supplementation improves selected physical performance parameters, but not cardiovascular responses in healthy older women: a randomized clinical trial
Aging is associated with declines in muscle strength and functional capacity, driven by both neuromuscular and cardiovascular changes. In parallel, ageing is accompanied by reduced nitric oxide (NO) bioavailability and potentially a reduced efficacy of the nitrate (NO₃-)–nitrite (NO₂-)–NO pathway, a route for NO generation that supports vascular and skeletal-muscle function. Thus, the aim of the present study was to examine the effects of acute ingestion of 70 mL of nitrate-rich beetroot juice (6.4 mmol NO₃-) versus 70 mL placebo (0.04 mmol NO₃-) on strength and functional performance in healthy older women aged between 70-80 years. Nine healthy women (75.4±4.0 years) participated in this randomized, double-blind, placebo-controlled crossover experiment. Participants completed a physical performance test consisting of 6-m walk test, isometric handgrip strength (i.e., dominant and non-dominant hand), Timed Up-and-Go Test (TUG), and sit-to-stand (STS) muscle power test along with cardiovascular assessments (systolic/diastolic blood pressure, heart rate response and 6-minutes’ walk performance) after ingesting either 70 mL of beetroot juice (6.4 mmol NO₃-) or 70 mL of placebo (0.04 mmol NO₃-) in a randomized order. Beetroot juice ingestion resulted in significant improvement vs. placebo in isometric dominant handgrip strength (9.6%, p=0.046, ES=0.78), with no changes observed in the non-dominant hand (1.2%, p=0.553, ES=0.11). No differences were found in beetroot juice vs placebo for 6-m walk performance (−3.3%, p=0.306, ES=−0.19), TUG (−5.0%, p=0.225, ES=−0.17), STS leg extensor muscle power (5.0%, p=0.121, ES=−0.25), or 6-minute walk performance (−7.7%, p=0.110, ES=−0.48). Acute ingestion of nitrate-rich beetroot juice led to increased isometric handgrip strength in healthy older women, with no significant effects observed in functional performance (TUG), lower limb STS power, cardiovascular responses, or short-term (6-m) as well as long-term (6-min) walking performance compared with placebo conditions.
Acute and chronic kidney injury following COVID-19 infection and vaccination: a narrative review
This narrative review examines acute and chronic kidney injury following COVID-19 infection and vaccination, discussing the mechanism of SARS-CoV-2 entry into host cells through the ACE2 receptor - highly expressed in renal tissues - facilitating the viral invasion. Viral RNA has been detected in the urine of patients infected with SARS-CoV-2, suggesting direct renal involvement. The incidence of Acute Kidney Injuriy (AKI) among hospitalized COVID-19 patients was particularly higher in the early stages of the pandemic and largely varied by 29%-46%, depending on population studied and COVID-19 wave. Pathological findings include Acute Tubular Injury (ATI), collapsing glomerulopathy, and focal segmental glomerulosclerosis. Major risk factors for AKI comprise older age, male sex, diabetes, hypertension, cardiovascular disease, and preexisting Chronic Kidney Disease (CKD). AKI significantly increases mortality of COVID-19 patients, particularly in advanced stages of renal failure. CKD is also associated with severe COVID-19 outcomes, including increased hospitalization, intensive care admission, and mortality. Patients with CKD show a dose-dependent relationship between disease stage and adverse patient outcomes. This review further addresses renal complications following COVID-19 vaccination, which, although rare, encompass various immune-mediated glomerular diseases. Minimal Change Disease (MCD) is most frequently reported after COVID-19 vaccination, followed by IgA nephropathy, membranous nephropathy, anti-glomerular basement membrane (anti-GBM) nephritis, and ANCA-associated vasculitis. These conditions commonly present with hematuria, proteinuria, or nephrotic syndrome, and many respond to corticosteroid or immunosuppressive therapy. Other less frequent renal complications include thrombotic microangiopathy, acute tubulointerstitial nephritis, and IgG4-related nephritis. In conclusion, both COVID-19 infection and vaccination can be associated with a spectrum of renal manifestations ranging from AKI to CKD and immune-mediated glomerulopathies. Awareness, early detection, and multidisciplinary management are essential to reduce renal morbidity and improve patient outcomes.
Risk factors and critical thresholds for glucocorticoid-induced ocular adverse reactions in children: implications for muscle and neuromuscular disease management
Systemic Glucocorticoid (GC) therapy is integral to managing various paediatric conditions, including inflammatory muscle and neuromuscular diseases. However, prolonged GC use in children poses a significant risk for severe ocular Adverse Drug Reactions (ADRs), notably Posterior Subcapsular Cataract (PSCC) and Steroid-Induced Ocular Hypertension (SIOH). These ADRs can lead to irreversible visual impairment, creating a substantial burden, particularly for paediatric patients already managing chronic conditions. Identifying specific risk factors and precise exposure thresholds is crucial for optimizing GC treatment and guiding timely ophthalmic monitoring. This retrospective case–control study analyzed data from 305 children with kidney disease, who received oral or intravenous GCs between December 2020 and December 2022. Participants were grouped into PSCC, SIOH, or control (no ocular ADRs). Logistic regression assessed risk factors, and Receiver Operating Characteristic (ROC) analysis determined critical thresholds for cumulative dose and duration. Results indicated seven significant risk factors for PSCC: younger age, longer GC duration, hypertension, growth retardation, vitamin D deficiency, immunosuppressant use, and oral methylprednisolone. Critical thresholds for cataract onset were identified as GC treatment duration >5.6 months or a cumulative dose >5,888 mg. For SIOH, higher daily GC dosage (>0.59 mg/kg/day), oral methylprednisolone, and growth retardation were significant risk factors. These findings elucidate key risk factors and establish quantitative GC exposure thresholds for ocular ADRs in children. They provide evidence-based guidance for ophthalmic screening and intervention timing, particularly relevant for paediatric patients with muscle and neuromuscular diseases requiring long-term GC therapy. Proactive monitoring and tailored GC management are essential to mitigate vision-threatening complications and improve long-term patient outcomes.
Beyond biomechanics: a narrative review on the cognitive, emotional, and metacognitive mechanisms of exercise in chronic pain
Chronic pain is increasingly recognized as a multidimensional condition sustained by interacting cognitive, behavioral, emotional, attentional, and metacognitive processes. Within this framework, physical exercise has gained relevance as a non-pharmacological intervention that may influence psychological mechanisms involved in pain persistence and disability. This narrative review provides an updated synthesis of recent evidence on the effects and mechanisms of physical exercise on cognitive, behavioral, emotional, and metacognitive dimensions of chronic pain. A structured literature search was conducted across PubMed/MEDLINE, Scopus and Web of Science, focusing on peer-reviewed studies published between January 2016 and December 2025. Evidence was synthesized narratively across mechanistic domains, integrating findings from rehabilitation science and clinical psychology. The literature suggests that exercise may act as an active experiential intervention capable of targeting several psychosocial factors in chronic pain. Across studies, exercise was associated with reductions in catastrophizing and maladaptive pain beliefs, improvements in pain self-efficacy, and attenuation of kinesiophobia. Exercise may also modulate attentional bias and hypervigilance, and psychologically informed exercise approaches appear particularly relevant for metacognitive processes, including reduced rumination, increased decentering, and improved non-reactive awareness of bodily sensations. In parallel, exercise shows beneficial effects on emotional aspects of chronic pain, likely mediated by both neurobiological and psychosocial mechanisms pathways. Overall, physical exercise should be conceptualized as a comprehensive intervention capable of acting on cognitive, behavioral, emotional, and metacognitive processes that sustain chronic pain. Future research should clarify modality-specific mechanisms, identify moderators of response and incorporate process-based outcomes to support personalized exercise prescriptions.
Handgrip strength in young recreational skiers: Is it related to the risk of falling?
Handgrip Strength (HGS) is correlated with general health and fitness throughout the lifespan. However, the relationship between HGS and the frequency of sports accidents is not well understood. The aim of the present survey was twofold. Firstly, to collect HGS data from young skiers and compare them with existing normative values; secondly, to investigate whether the frequency of falls during a day of skiing is correlated with HGS. Data on HGS and falls were collected from 251 recreational downhill skiers, aged 6–25 years, on the day of recording. The results demonstrate the representative progression of HGS in boys and girls during growth and early adulthood. Additionally, the findings indicate that in young female skiers up to the age of 25 years, HGS is negatively correlated with the frequency of falls, which may be related to the generally lower muscle strength in females. Regular physical activity, including muscle strengthening and balance exercises, is crucial for preventing falls in skiers.
Changes in physical performance with aging in master athletes and in the general population: an update
One of the fundamental biological processes underlying aging is the decline in physical performance. Sarcopenia, dynapenia, chronic inflammation, and other factors including the loss of motor units largely account for this decline. Master athletes—individuals who train and compete well beyond early adulthood—represent a valuable model for studying healthy aging. In the general population, physical performance follows a characteristic lifespan trajectory, increasing from childhood to a peak in early adulthood and progressively declining with aging due to a reduction in muscle mass and quality, and in multisystem physiological functions. The seminal studies by Gava and colleagues on master athletes indicate that, under ideal conditions of being disease-free or injury-free, performance loss of these athletes follows an attenuated, linear fashion from early adulthood into advanced age. While lifelong training cannot halt age-related physiological deterioration, it can attenuate the rate of functional decline in cardiovascular, neurocognitive, and musculoskeletal functions. From a sex-based perspective, males demonstrate an absolute advantage in power and strength compared to females, whereas differences in endurance performance are smaller. With age, the physical performance gap between the two sexes tends to narrow, particularly in endurance disciplines. Overall, the master athlete model supports the concept that aging-related performance decay follows predictable biological rules, while its rate remains highly modifiable through sustained physical activity. These insights have important implications for exercise prescription, preventive strategies, and healthy aging
Infection prevention in total knee arthroplasty: current concepts, controversies, and a clinical algorithm
Periprosthetic Joint Infection (PJI) remains one of the most serious and challenging complications following Total Knee Arthroplasty (TKA), with a substantial impact on patient outcomes and healthcare systems. Despite advances in surgical techniques and perioperative management, the incidence of PJI has not significantly declined, highlighting the need for more effective preventive strategies. This review aims to provide a comprehensive and clinically oriented overview of current infection prevention strategies in TKA, with a particular focus on risk stratification, perioperative optimization, and the development of a practical clinical algorithm to guide decision-making. A narrative review of the literature was conducted using major medical databases to identify relevant studies on PJI prevention in TKA. Priority was given to recent systematic reviews, meta-analyses, and high-quality clinical studies addressing patient-related risk factors, intraoperative measures, and postoperative management. PJI is a multifactorial complication influenced by the interaction between patient-related, surgical, and postoperative factors. Modifiable risk factors such as obesity, diabetes, malnutrition, and smoking play a pivotal role and should be optimized preoperatively. Intraoperative strategies, including strict aseptic technique, appropriate antibiotic prophylaxis, and minimization of operative time, are essential to reduce contamination risk. Postoperative measures focusing on wound management, hematoma prevention, and early detection of complications further contribute to infection prevention. Several aspects remain controversial, including the role of laminar airflow systems, antibiotic-loaded bone cement in primary TKA, and optimal decolonization strategies for Staphylococcus aureus. Effective prevention of PJI requires a structured, multimodal approach integrating all phases of care. Risk stratification enables personalized preventive strategies and improved perioperative planning. The clinical algorithm proposed in this review provides a practical framework to support decision-making and may help standardize infection prevention pathways. Future research should focus on validating risk-based approaches and emerging technologies to further reduce the burden of PJI in TKA.
34 | Hyaluronan enhances proliferation and myogenic differentiation of C2C12 murine myoblasts under inflammatory stress
Hyaluronan (HA) is a non-sulfated glycosaminoglycan, widely used for medical and pharmaceutical applications including tissue muscle repair. A recent study demonstrates that HA activates muscle stem cells to repair damaged muscle. When muscle damage occurs, stem cells start producing and coating themselves with hyaluronic acid, driven by demethylase JMJD3, which allows muscle stem cell adaptation to inflammation and the initiation of muscle repair (Nakka et al., 2022). The aim of this work is to characterize the effect of Multifractionated - HA on myoblast rescue under inflammatory conditions, using C2C12 murine muscle cell proliferation and differentiation. In this study, we investigated the potential of a HA mixture (2-1000 KDa, 1 mg/mL, Regenflex TeM, Regenyal Laboratories SRL) in increasing the proliferative and myogenic capacity of myoblasts in the presence or absence of pro-inflammatory agents (IL-1β, LPS), known to impair proliferation. Results revealed that TeM significantly improved reparative mechanisms and exerts a strong pro-proliferative effect, enhancing wound healing within 24 hours post-injury, even in the presence of inflammatory conditions. Additionally, we evaluated the myogenic potential of C2C12 cells treated with TeM by analyzing the gene and protein expression of key myogenic markers, including IGF-1, MyoD, Myogenin, Mrf4, myogenin, and MHC-1, even under inflammatory conditions. Moreover, TeM treatment upregulated myogenic biomarkers, suggesting a positive impact on differentiation pathways. Regenflex T&M has been shown to increase the repair process in injured myoblast monolayers, to restore healing capacity under otherwise limiting conditions and to stimulate the muscle differentation process. This data streghtens the rationale for using infiltrative intramuscular T&M to promote the repair of injured muscles in clinical settings.
Abstracts of the 22nd Meeting of the Interuniversity Institute of Myology
Copyright (c) 2026 The Author(s) https://creativecommons.org/licenses/by-nc/4.0
43 | Synergistic negative effects of advanced glycation end products and tumor-conditioned medium on C2C12 myotubes and the protective activity of Vaccinium macrocarpon
Cancer cachexia (CC) is a complex and multifactorial syndrome characterized by progressive loss of skeletal muscle mass that could be worsened by unhealthy dietary habits, such as the consumption of a Western diet (WD), which supplies high quantities of advanced glycation end products (AGEs), non-enzymatic adducts formed through the reaction of reducing sugars with cellular macromolecules that can can alter the structure and function of the latter. An excessive accumulation of AGEs has been observed in both the plasma and tumor microenvironment of cancer patients, where it correlates with the severity of muscle wasting. In recent years, natural compounds have emerged as promising candidates for treatment due to their diverse bioactive properties. Among these, Vaccinium macrocarpon (VM) has been identified through preliminary ad hoc screening as a potent inhibitor of AGE formation and activity, suggesting its potential as a therapeutic agent against AGE-induced muscle wasting. Since the degeneration of the neuromuscular junction (NMJ) may contribute to CC-related muscle wasting, we explored the role of AGEs in the dismantling on NMJ and the therapeutic potential effect of VM, with a particular focus the effects on AChR formation and clustering, a well-established in vitro model for evaluating NMJ development and function. Our data indicate that AGEs cause the worsening of atrophy of C2C12 myotubes and a reduction of AChR clustering (an in vitro model of NMJ formation) induced by the treatment of cells with Lewis lung carcinoma conditioned medium (LLC-CM). Co-treatment with VM rescued myotube diameter and AChR clustering, suggesting its potential to mitigate the detrimental effects on skeletal muscle of WD associated with CC.
28 | Impairment of skeletal muscle force and mitochondrial respiratory capacity precedes muscle mass loss in an aged murine model of diet-induced obesity
Sarcopenic obesity (SO) is characterized by the coexistence of obesity and sarcopenia, conditions that synergistically exacerbate frailty, particularly in the elderly population. However, the mechanisms underlying the development of sarcopenia within the context of obesity remain poorly understood. It is well established that mitochondrial respiratory capacity in skeletal muscle declines with age, and it is plausible that this decline contributes to the pathogenesis of SO. In this study, using a diet-induced murine model of obesity, we investigated the progression of sarcopenia in obesity during aging, with a particular focus on skeletal muscle mitochondria. We analyzed 12-month-old female C57BL/6 mice fed either a high-fat diet (HFD; 60% of calories from fat) or a standard diet (SD; 10% of calories from fat) over a seven-month period. We monitored body weight and composition in vivo, muscle volume, muscle strength and endurance. At 1, 3, and 7 months of diet, muscles were collected from both HFD- and SD-fed mice for histological and molecular ex vivo analyses. We evaluated respiration, number, area, diameter of mitochindria and mitochondrial network. From the first month of HFD feeding, mice exhibited increased body weight and fat accumulation. Grip strength, used as an indicator of muscle function, was significantly reduced in HFD-fed mice after one month, with further decline observed at three months. By seven months, histological analysis revealed a marked reduction in myofiber cross-sectional area compared to SD-fed controls. Notably, mitochondrial respiratory capacity and mitochondrial number were initially elevated after one month of HFD but showed a significant decline by three months. These results suggest that early metabolic alterations in mitochondrial function contribute to the onset and progression of sarcopenia in the context of obesity.
45 | Interleukin-4 prevents cancer-induced muscle wasting and promotes exercise-like metabolic adaptations
Cancer cachexia is a complex syndrome characterized by body weight loss, muscle wasting, mitochondrial abnormalities and energy metabolism alterations. Previous findings showed that treating tumor-bearing (TB) mice with interleukin-4 (IL4) improved muscle fitness and myogenesis. IL4 and IL4 receptor were also found to be upregulated in the skeletal muscle after exercise. The present study aimed to investigate whether IL4 administration improves skeletal muscle energy metabolism in cancer hosts. Ten-weeks-old Balb/c male mice were inoculated with 5x105 C26 colon carcinoma cells and sacrificed at days 10 or 15. Daily IL4 treatment (66.5 μg/kg) was performed by intraperitoneal injection. The gastrocnemius muscle was used to assess mitochondrial proteins (western blot) and respiration (high-resolution respirometry, Oroboros Instruments), and to perform metabolomic and lipidomic profiling (HPLC-coupled mass spectrometry). Treatment with IL4 prevented loss of body weight, muscle mass and muscle strength in day 15 TB mice. The protein levels of OXPHOS complexes II and III, cytochrome c and PGC-1α in the skeletal muscle were significantly increased in treated vs untreated TB mice. Similar trends were found for TOMM20, while trends to decrease were found for BNIP3. OXPHOS complex II activity was significantly increased in treated vs untreated TB mice. Tumor- and IL4-driven modulations in skeletal muscle metabolomic profile were observed, involving many energy-associated metabolites. Similarly, the skeletal muscle lipidome was altered by tumor burden and was partially restored by IL4. IL4 treatment improves energy metabolism in the skeletal muscle of TB mice. This effect relies on the improvement of mitochondrial homeostasis and the increased availability and capacity of mitochondrial OXPHOS complexes, suggesting that IL4 may act as an exercise mimetic. Further efforts to unravel the mechanism underlying IL4 efficacy are required.
26 | Validation of a new multi-target antioxidant treatment for the therapy of Duchenne muscular dystrophy
Currently, there is no general cure for Duchenne muscular dystrophy (DMD); standard therapy with corticosteroids only slows the progression of the disease, while often causing significant side effects. Therefore, there is a clear need for new, mutation-independent effective therapeutics with favourable side effect profiles. In DMD, dystrophin deficiency triggers a series of pathophysiological events, e.g., calcium dysregulation, mitochondrial disfunction, oxidative stress, inflammatory and immune responses, and impaired muscle regeneration. Experimental therapies in the past usually focused on only one of these aspects at the time, limiting the efficacy of these approaches. A therapeutic strategy capable of simultaneously acting on multiple DMD pathophysiological features would therefore be highly desirable. To this aim, we identified the key transcriptional regulator NRF2 as a promising pharmacological target, which can be specifically activated by the small molecule Nacetylcysteine ethyl ester (NACET). Here we report the initial results obtained by treating young mdx mice with NACET from 6 to 10 weeks of age. So far, we found that the highest dosage led to a significant improvement of normalized force in hindlimb muscles and to a decrease in the percentage of necrotic fibers, in the absence of any adverse side effects.
16 | Immunoproteasome inhibition positively impacts the gut-muscle axis in Duchenne muscular dystrophy
Duchenne muscular dystrophy (DMD), a fatal muscle-wasting disorder resulting from mutations in the DMDgene, is often compounded by secondary non-muscle complications. A key factor in DMD progression is chronic inflammation, triggered by the release of Danger Associated Molecular Patterns (DAMPs) following myofiber necrosis, which attracts proinflammatory cells to dystrophic muscle. Notably, gastrointestinal (GI) dysfunction, common in DMD patients, can also contribute to systemic inflammation via the gut microbiota. The immunoproteasome (IP) plays a crucial role in regulating antigen presentation, cytokine production, and immune cell responses, and acts as a first line of defense in the gut by modulating the NF-κB pathway. Our prior research demonstrated that IP inhibition alleviates molecular, histological, and functional deficits in mdx mice, a well-established DMD model. Here, we investigated the impact of IP inhibition on the gut-muscle axis in these mice. Treatment with the IP inhibitor ONX-0914 increased the presence of anti-inflammatory CD206+ M2 macrophages in both the colon and muscle of mdx mice, while partially reduced pro-inflammatory CD68+ M1 cells. This treatment also improved muscular mitochondrial activity, enhanced muscle performance, and decreased serum creatine phosphokinase (CPK) levels and fibrosis. Utilizing microbiota perturbation models, including broad-spectrum antibiotics and fecal microbiota transplantation from IP-inhibited mdx mice, we demonstrated that ONX-0914's beneficial effects on muscle function are mediated through microbiotadependent mechanisms. Our findings contribute to understanding the role of IP inhibition as a potential therapeutic strategy for modulating the dystrophic gut-muscle axis, suggesting new avenues for microbiota-targeted therapies.
37 | Characterization of novel cancer cachexia preclinical models implementing chemotherapy and surgery
Cancer cachexia (CC) is a multifactorial syndrome characterized by weight loss, metabolic alterations, and muscle wasting. Although the C26-mouse model is widely used to study CC, its rapid tumor growth and lack of metastasis fail to recapitulate cancer patient’s trajectory. In colorectal cancer, standard treatment includes chemotherapy and resection of the primary tumor, often followed by tumor relapse and/or metastasis. This study aims to characterize novel C26-based experimental protocols to investigate tissue-specific metabolic changes. Three treatment protocols were evaluated in 3-month-old female BALB/c mice subcutaneously inoculated with 5×10⁵ colon carcinoma cells (C26). Each protocol combined FOLFOX chemotherapy with surgical resection: (1) two doses pre-resection (C26+2F), (2) two doses pre- and post-resection (C26+4F), and (3) a single dose pre-resection (C26+F). Tumors were surgically excised 4 days post-chemotherapy. A group of control mice was included in each experiment. At sacrifice, muscles and tissues were collected for biochemical, histological, and metabolomic analyses. Tumor-bearing mice showed signs of cachexia in all experiments, with increasing severity according to chemotherapy duration. Few mice showed no evidence of primary tumor relapse, fully recovering their body weight and muscle mass. Others progressed to lung metastases, indicating that curative interventions lead to distinct trajectories even in syngeneic mice. Metabolomics analysis in gastrocnemius and liver revealed distinct metabolic profiles of control and tumor-bearing mice, with the no relapsing mice clustering closer to the controls. Muscle and liver exhibit distinct metabolic alterations, showing signs of potential recovery depending on treatment strategy and early intervention. Further investigations are required to identify affected metabolic pathways associated with either bad prognosis or cancer survivorship and recovery.
01 | Investigating skeletal muscle-endothelial cell crosstalk in plasticity and disease
The maintenance of skeletal muscle mass and function is a major clinical challenge in aging, cancer, muscular dystrophies, and related conditions. While much attention over the past two decades has focused on intracellular pathways like Akt-mTORC1, the roles of muscle perfusion and paracrine signaling between muscle fibers and endothelial cells (ECs) remain poorly understood. Emerging evidence suggests a bidirectional crosstalk between these cell types. However, in vivo tools to study this interaction with cell-type specificity and temporal control are still limited. To address this, we developed an EC-specific MetRS mouse model expressing a mutant methionyl-tRNA synthetase (MetRS^L274G) under the tamoxifen-inducible Cdh5-CreERT2 promoter. This allows temporally controlled, EC-specific incorporation of the methionine analog azidonorleucine (ANL) into nascent proteins in vivo. Administered via drinking water, ANL selectively labels EC proteins within defined windows. Labeled proteins can be visualized and quantified using click chemistry–based FUNCAT or enriched by immunoprecipitation for downstream mass spectrometry–based proteomic analysis. Combined with tissue clearing and light sheet microscopy, this enables 3D mapping of endothelial protein synthesis across skeletal muscle. To validate the model, we induced angiogenesis via unilateral AAV-VEGF overexpression (2×10¹⁰ U/muscle) in the gastrocnemius muscle, using the contralateral limb as control. Our preliminary results demonstrate successful and specific endothelial protein labeling with clear spatial distribution across muscle tissue. Interestingly, VEGF-induced angiogenesis correlates with increased muscle mass, fiber size, and protein synthesis. We are now using the model to explore how controlled muscle growth or atrophy influences ECs, and how primary EC changes affect muscle physiology. This work aims to provide new insights into muscle–vascular communication relevant to muscle health and disease.
32 | Trifolium pratense extract mitigates skeletal muscle atrophy: a nutraceutical approach for the prevention of sarcopenia
Sarcopenia, the age-related loss of muscle mass and function, increases the risk of frailty, fractures, and loss of independence, ultimately reducing quality of life and contributing to higher mortality. With life expectancy rising in Western countries, sarcopenia represents a growing yet unresolved public health challenge, as no pharmacological treatments are currently available. Recent interest in medicinal plants as potential therapeutic agents has highlighted their ability to counteract various pathological conditions, including muscle atrophy. This study explores the protective effects of native Mediterranean on skeletal muscle cells exposed to atrophic stimuli that mimic sarcopenia and disuse (e.g., dexamethasone, starvation). Additionally, we assess whether controlled cultivation protocols can influence the bioactive metabolite content of these plants, potentially enhancing their anti-sarcopenic properties. Among the tested species, hydroalcoholic extract of Trifolium pratense demonstrates promising anti-atrophic effects, effectively preserving C2C12 myotube diameter against TNF- α/IFN-γ-induced muscle wasting. However, its protective efficacy varies depending on cultivation conditions, with some enhancing its bioactivity while others showing no significant effect. Furthermore, Trifolium pratense completely abolishes TNF-α/IFN-γ-induced reactive oxygen species production, underscoring its antioxidant properties. These findings suggest that Trifolium pratense could serve as a promising nutraceutical intervention to counteract muscle atrophy associated with aging. Moreover, optimizing its cultivation conditions may enhance its therapeutic potential, paving the way for targeted agronomic strategies to maximize its bioactivity. By integrating plant-based interventions into sarcopenia management, this study contributes to the development of accessible, nonpharmacological strategies aimed at improving muscle health and quality of life in aging populations.
12 | In vitro study emulating muscle response to stimuli exploiting 3D tissue model
Understanding muscle responses to external stimuli is critical for advancing therapeutic strategies and vaccine delivery, yet human-based models remain limited. To emulate muscle responses to immunological and delivery-related stimuli in vitro, we developed a human 3D muscle-like tissue model coupled with peripheral blood mononuclear cells (PBMCs). Upon exposure to immunostimulatory agents, the system supported the recruitment and modulation of antigen-presenting cells, including monocytes and macrophages, while stromal elements promoted dendritic cell expansion and activation. PBMCs exhibited an early innate immune signature (encompassing IL-15, IL-6, TNF-α, IFN-γ, IP-10, MCP-1, and IFN-inducible genes) reflecting immune activation profiles relevant for protective responses. Muscle-resident cells fostered an immunostimulatory environment, while the stromal compartment enhanced IFN-related mediator expression, demonstrating the platform’s capability to investigate muscle-immune system interactions under controlled conditions. To further explore muscle responses to delivery stimuli, we assessed the internalization and modulatory effects of solid lipid nanoparticles (SLN) and poly(lactic-co-glycolic acid) (PLGA) nanoparticles within the 3D muscle constructs. Both nanoparticle types were progressively internalized by myofibers, exhibiting distinct localization and accumulation near nuclei. Additionally, we tested the capability of SLN and PLGA nanoparticles to modulate inflammatory cytokines (IL-1β, IL-6, TNF-α) under inflammatory stimulation, indicating their potential for controlled delivery and local immune modulation within muscle tissue. In summary, this human 3D muscle-based in vitro model effectively emulates muscle responses to immunological and delivery stimuli, offering a versatile platform to study muscle immunobiology and evaluate advanced delivery strategies, while reducing the reliance on animal models in preclinical research.
03 | The muscle out of breath? Limitations and adaptations in chronic diseases
Skeletal muscle demonstrates remarkable plasticity in response to physiological stimuli such as exercise and hypoxia. Chronic diseases like chronic obstructive pulmonary disease (COPD), chronic heart failure (CHF), and obstructive sleep apnea syndrome (OSAS) result in chronic or intermittent hypoxia, impairing oxygen delivery to muscle tissue. Sickle cell disease (SCD), a genetic hemoglobinopathy, represents a unique model characterized by a complex hypoxic environnement (chronic, acute, and intermittent) stemming from severe chronic hemolytic anemia and recurrent vaso-occlusive crises. These hypoxic conditions induce profound alterations in skeletal muscle, mostly including fiber-type switching, atrophy, reduced capillarization, and impaired oxidative metabolism. The extent of these changes appears to depend on the nature and severity of hypoxia, as well as the degree of physical inactivity. In COPD and CHF, exercise training is well established to mitigate muscle dysfunction. Conversely, in SCD, physical activity was long considered risky due to fears of provoking vaso-occlusive events. However, our research group conducted a randomized controlled trial demonstrating that moderate-intensity endurance training is not only safe in individuals with SCD, but also enhances physical capacity and attenuates muscle alterations. These findings pave the way for novel therapeutic strategies based on exercise to improve muscle structure and quality of life in this population. In contrast, the effects of exercise training on skeletal muscle structure in patients with OSAS remain unexplored, highlighting a critical gap in the current understanding of muscle adaptation in hypoxia-related disorders.
40 | Role of glucocorticoids in cancer cachexia: focus on lipocalin 2 and iron homeostasis
Cachexia is a severe wasting syndrome characterized by profound skeletal muscle atrophy that significantly contributes to mortality across various diseases, including cancer. However, its underlying mechanisms remain poorly understood. We previously identified major alterations of iron metabolism in the skeletal muscle of cachectic tumor-bearing mice and in particular a decrease in mitochondrial iron content, resulting in mitochondrial dysfunction. Lipocalin 2 (LCN2), an iron-chelating protein, has been identified as one of the most upregulated genes in various models of skeletal muscle atrophy. Being known the role of glucocorticoids in muscle wasting and that LCN2 is a transcriptional target of the glucocorticoid receptor (GR), we hypothesized its involvement in the dysregulation of iron metabolism during muscle wasting. Firstly, we confirmed the increased expression of LCN2 in the skeletal muscle of both C26 tumor-bearing mice and dexamethasone-treated mice, recapitulating the cachectic phenotype. Consistently, in vitro LCN2 overexpression induced a significant reduction in C2C12 myotube diameter and, oppositely, its silencing was sufficient to prevent dexamethasone-induced atrophy. Moreover, iron supplementation prevented glucocorticoid-induced atrophy in vitro and mitigated the cachectic phenotype in our two mouse models. Importantly, local expression of LCN2 promoted markers of iron deprivation in vivo. These results suggest that iron metabolism acts as a key contributor to skeletal muscle atrophy. Our findings shed light on both cancer and glucocorticoid-induced skeletal muscle wasting mechanisms, highlighting LCN2 as a potential therapeutic target for muscle wasting diseases.
Welcome to the 22nd Meeting of the Interuniversity Institute of Myology!
The 22nd Meeting of the Interuniversity Institute of Myology (IIM) took place in Assisi, Italy, from September 11 to 14, 2025, welcoming 106 participants from academia, industry, and patient organizations across Europe and the USA. The meeting provided an interdisciplinary forum for discussing muscle tissue from every perspective, combining scientific sessions with informal social events to foster networking. Early-career scientists played a central role, representing half of the attendees. Their involvement was supported by the High Training Course “Advanced Myology Update 2025,” jointly organized by the University of Perugia and IIM, which hosted 15 participants in lectures and roundtable discussions led by invited experts. The scientific program comprised six sessions, including 28 oral presentations and 36 posters. Four keynote lectures by Karyn Esser, Markus Rüegg, Giulio Cossu, and Roberta Sartori offered in-depth insights into muscle biology, neuromuscular disorders, and translational research. In line with IIM’s support for young scientists, presenters under 35 were eligible for Best Talk, Best Poster Blitz, and Best Poster awards. Winners were invited to join the IIM Young Committee, contributing to future meeting organization. Overall, the 22nd IIM Meeting confirmed its role as an inclusive and dynamic forum for the myology community. It successfully combined high-level science with opportunities for mentorship and collaboration, reinforcing IIM’s international relevance. We look forward to welcoming both long-standing members and new participants at the 23rd IIM Meeting in Assisi, September 9-12, 2026, for another engaging event full of science and shared enthusiasm.
27 | Polymeric nanoparticles as a favorable delivery system to enhance the anti-inflammatory action of n-palmitoylethanolamide, an endocannabinoid-like molecule
Chronic low-grade inflammation is linked to age-related muscle loss (sarcopenia), highlighting the potential of daily anti-inflammatory strategies. A growing body of evidence suggests that N-palmitoylethanolamide (PEA), a natural endocannabinoid-like compound with anti-inflammatory properties, could represent a safe therapeutic option to counteract sarcopenia. However, the low solubility of PEA significantly limits its systemic bioavailability. To overcome this limitation, our work was aimed to develop a nanoparticle-based delivery system intended to enhance its systemic availability. We compared two nanoparticle formulations: solid lipid nanoparticles (SLNs) and poly-(lactic-co-glycolic acid) (PLGA) polymeric nanoparticles. These delivery systems were evaluated in terms of cellular uptake, cytotoxicity, and impact on myogenic differentiation. Our results indicate that PLGA nanoparticles exhibit superior cellular internalization compared to SLNs, display lower cytotoxicity, and do not substantially interfere with the myogenic differentiation process in cell culture. Furthermore, PEA encapsulated within PLGA nanoparticles effectively activated its canonical target, peroxisome proliferator-activated receptor alpha (PPAR-α), and protected muscle cells from lipopolysaccharide (LPS)-induced cytotoxicity in vitro. These findings represent a critical step toward the preclinical evaluation of PEA as a potential therapeutic agent for counteracting sarcopenia, using murine models of age-related muscle wasting. Funding: European Union Next Generation EU” n. P2022LSW98 (PRIN-PNRR 2022 from Italian "Ministero dell'Università e della Ricerca".
35 | A novel 3D skeletal muscle model to study cancer cachexia
Cancer cachexia is a multisystem syndrome affecting cancer patients, characterized by progressive body weight loss, muscle wasting, increased mortality and poor quality of life. Skeletal muscle represents a main target of cancer cachexia, where tumor-derived inflammatory factors including IL-1, IL-6 and TNF-α trigger muscle degeneration through the activation of specific proteolytic pathways, ultimately causing a dramatic loss of muscle mass and function. Although several mouse models have been used to study cancer cachexia, the close interaction between tumor and muscle cells has been investigated primarily in 2D in vitro co-culture models. This approach revealed important molecular and metabolic dysfunctions of muscle cells when co-cultured with tumor cells. However, the use of 2D systems to study skeletal muscle differentiation/degeneration dynamics presents many limitations, including the lack of the characteristic spatial orientation of the myotubes. Here, we present a novel hydrogel-based 3D in vitro muscle model to study cancer cachexia. We generated bio-artificial muscle and extended the model by incorporating a co-culture with colorectal carcinoma cells, enabling direct cell–cell contact between cancer cells and myogenic cells. After differentiation in co-cultures with carcinoma cells, myoblasts showed a severe impairment of their myogenic potential. This model also allows to uncouple direct cellcell contacts while studying the effect of secreted factors by an indirect co-culture system. In addition, we found that the typical myofiber alignment observed in native muscle tissue is preserved in our constructs also in the presence of carcinoma cells. The ability to study cancer-related myogenic dysfunction in a soft 3D environment represents a novel tool to study cancer cachexia mechanisms and drug development, further reducing the need for animal experiments.
29 | RNA-based combinatorial multi-target therapy for the treatment of Duchenne muscular dystrophy
Duchenne muscular dystrophy (DMD) is a severe and fatal X-linked desease, caracterize by the lack of dystrophin leading to structural fragility of muscle fibers and promotes a pro-catabolic metabolic imbalance. To date, no curative treatment is available, partly due to the difficulty in restoring full dystrophin expression and developing non-immunogenic approaches. Reactivation of utrophin, a dystrophin’s paralogous protein naturally downregulated by microRNAs, has shown strong potential to compensate for dystrophin deficiency. The RNA_DMD project aims to address three major challenges that are 1) to reactivate utrophin expression to compensate for the loss of dystrophin, 2) to inhibit pro-catabolic pathways to promote muscle growth, 3) to develop non-immunogenic vectors for effective and tolerable combination therapy. We hypothesize that combination therapy could both restore muscle structural integrity and prevent or reverse muscle wasting and associated loss of function. To study aim 2, we used the bicistronic plasmid expressing green fluorescent protein (GFP) and bearing our shRNA against FOXO1/3 or MURF1caring by an adeno-associated virus with a muscle specific serotype (AAV-MYO). To date, we test the efficiency of our AAV-MYO-shRNA in wild-type (WT) mice by electroporation in tibialis anterior (TA) (1010 vg/muslce) and systemically (1012 vg/mouse). And then, we moved to 3-week-old D2-mdx mice (a severe model of DMD). Our preliminary data highlights in WT animals, that combined electroporation of AAV-MYOshMuRF1 and AAV-MYO-shFOXO1/3 in the tibialis anterior muscle leads to a reduction in the expression of MuRF1 (-24%; P=0.04), FOXO1 (-36%; P=0.002), and 3 (-39%; P=0.03) genes, associated with muscle hypertrophy described by a +30% increase in CSA (P=0.03) of TA. In 3-week-old D2-mdx mice separate intramuscular injection of AAV-MYO-shMuRF1 and AAVMYO- shFOXO1/3 led to a reduction after 4 weeks of MuRF1 transcript (-45%; P<0.001) and FOXO1 respectively (-27%; P=0.04).
05 | Stimulation of muscle stem cells using side-streams from animal and plant production
Current estimates predict that global food production needs to increase and adapt in response to population growth and dietary changes. Better utilization of animal and plantbased side-streams from food production will be necessary to achieve a circular economy while maintaining the nutritional needs of the global population. Muscle health and regenerative properties are important during aging, diabetes, and normal skeletal muscle homeostasis, and are highly affected by nutrient intake. Muscle stem cells, also known as satellite cells, play a key role in muscle repair and regeneration. Metabolic pathways involving glucose utilization are essential for the proper functioning of these stem cells. Muscle stem cells and their regenerative properties have been shown to be stimulated by small peptides and chemical compounds like phytochemicals, influencing proliferation, glucose uptake, and protein synthesis. In this study, we explore the potential of hydrolysates from side-streams of the chicken industry and phytochemical-rich extracts from plant sidestreams in glucose uptake and muscle regeneration. Our data showed that peptides generated from chicken hydrolysates and extracts from cauliflower, kale, and apple stimulated the proliferation of bovine muscle cells using the Incucyte live cell system. Further analysis revealed that chicken hydrolysate increased glucose uptake and protein synthesis in both bovine and rat muscle cells through regulation of the MEK and ERK signaling pathways. In conclusion, we observe bioactive compounds in rest-raw materials from both plants and chicken that can be promising nutraceutical supplements for muscle health.
44 | Targeted delivery of mir-486 to skeletal muscle: efficacy, tropism, and integrin dependency of MYOAAV2A vs. AAV9
Adeno-Associated Viruses (AAVs) are emerging as a promising tool for gene therapy as a way to modulate gene expression in specific tissues. Of particular interest to us is the application of AAV-based approach for treating congenital myopathies. The use of AAV in the muscle is complicated by the abundance and large distribution of this tissue in the body, requiring high dosages of the gene-delivery vector with higher risks of side effects, mostly related to liver toxicity. To address this challenge, we designed and selected a strong expression cassette for miR-486, known to ameliorate myopathic symptoms, through in vitro screening, and loaded it in the recently developed myotropic AAV virus MyoAAV2A to enhance skeletal muscle transduction and targeting. This delivery system achieves a high transduction and expression of miR-486 in mouse striated muscle while de-targeting the liver and the lungs. The higher muscle trophism and efficiency of MYOAAV2A compared to its parental serotype AAV9 depend on the presence of the integrin recognition motif, RGD, in the hypervariable region VIII of the virus capsid, possibly making its infection efficiency dependent on the presence of RGD-binding integrins (particularly αVβ6) in the sarcolemma. Our current understanding of how αVβ6 expression varies with age, muscle type, and in non-muscle organs is still limited, yet this knowledge could be valuable for designing MYOAAV-based therapeutic strategies. Here, we analysed the correlation between the transduction efficiency of MyoAAV2A and αVβ6 expression across various muscles, organs and mouse ages, comparing it with AAV9 as an integrin-independent control serotype.
46 | Investigating the impact of the tumor secretome on in vitro human skeletal and cardiac muscle models of cancer-induced cachexia
Cancer cachexia is a complex and poorly diagnosed metabolic syndrome primarily characterized by depletion of skeletal muscle and adipose tissue. Although cachexia impacts multiple organs, including the heart, research has remained focused on skeletal muscle, with no robust in vitro model of cardiac cachexia available. Furthermore, current knowledge regarding cancer cachexia is mainly derived from animal studies, which lack direct translatability to human physiology. Therefore, this project aims to bridge these gaps by integrating an established murine model of cancer cachexia with advanced human in vitro systems and, ultimately, patient-derived tissue explants to investigate the effects of the tumor secretome on skeletal and cardiac muscles. Specifically, we developed fully human in vitro models by indirectly exposing immortalized myoblasts to the secretome of human colon carcinoma cells (HCT116) using transwell-based co-cultures, which led to reduced expression of mature myogenic markers and formation of smaller myotubes. Applying the same approach to human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes, we observed impaired cardiac differentiation and maturation as well as reduced cardiomyocyte contractile function. Experiments with hiPSC-derived cardiac organoids, which offer increased complexity and greater physiological relevance, are ongoing. In parallel, since we have observed more small extracellular vesicles (sEVs) containing stress-related proteins in cachectic cancer patients compared to weight stable patients, we have begun testing the sEV inhibitor Manumycin A in our cell systems. These experiments showed reduced sEV secretion, and we plan to assess whether sEV inhibition can mitigate cachectic phenotypes in our skeletal and cardiac models. By leveraging state-of-the-art human models and sEV-modulating interventions, this research will provide crucial insights into cachexia-induced alterations in skeletal and cardiac muscles.