PN 23.16.01.02

Brief presentation of the project

Muscle sarcopenia/cachexia is defined as a loss of muscle mass, decreased muscle strength and striated muscle functionality, leading to disability and diminished quality of life and even progression of cognitive decline. In Parkinson's disease (PD), for example, 40% of patients suffer from this syndrome and 11% reach a severe form. The relationship between muscle wasting and neurological deficit has not yet been demonstrated. Recent studies have shown that there is an amelioration of symptoms in some neurodegenerative diseases after low/moderate intensity physical exercise that induces favorable changes in striated muscle tissue. In addition, in age-related cerebral atrophy, physical exercise proves effective, improving cerebral perfusion, metabolic functions, mitochondrial biogenesis, and reducing oxidative stress and neuroinflammation. Our experimental in vitro and in vivo models of BP as "proof of concept", will allow the extrapolation of the results obtained to the human pathology of BP, possibly also with reference to other neurodegenerative diseases with muscle damage. Skeletal muscle is an endocrine organ that produces and releases myokines (muscle-specific cytokines), which have been shown to exert specific endocrine effects on other organs, including the brain. The collaboration with the Colentina Clinical Hospital gives us access to biological samples – blood and CSF, from BP patients. Our study is original and novel, both nationally and internationally. The panel of muscle biomarkers that we aim to establish to assess the communication between the muscle system and the central nervous system on the muscle-brain axis includes both myokines and epigenetic markers, especially muscle miRNAs, but also proteins involved in epigenetic mechanisms and oxidative stress (DNMTs, SIRTs).

The aim of the project: to establish a panel of circulating muscle biomarkers which are modified in BP with muscle cachexia, to evaluate the prognosis of the disease and as possible therapeutic targets. This approach will allow us to identify some molecules that could be intercepted or supplemented in order to delay or prevent the disruption of the cellular mechanisms underlying the damage to the two tissues, on the brain-muscle axis. For this purpose we use "proof of concept" experimental models in vitro and in vivo. In the in vitro experimental model we use human neuroblasts SH-SY5Y cell line differentiated into dopaminergic neurons in which the pathological accumulation of alpha-synuclein/phospho-alpha-synuclein is induced by treatment with rotenone or MPP+. These cells are co-cultured with normal human skeletal muscle cells differentiated from the human myoblast line HSkMC. Quantitatively/qualitatively modified biomarkers are detected both at the cellular level (for both cell types) and in the culture medium, as a result of the interaction, through soluble molecules, between the two cell populations. Another experimental in vitro model is that of premature muscle aging induced with antisense oligonucleotides (ASOs) in murine C2C12 cells to induce alternative splicing in the LMNA gene leading to the loss of 150 bases from the end of exon 11. A quantitative/qualitative analysis of modified biomarkers is done at the cellular level and also it is tested the influence of this phenotype on neuronal cells. For the in vivo experimental model we use transgenic mice for human Parkinson's disease: B6;C3-Tg(Pmp-SNCA* A52T)83Vle/J. We are looking for a significant weight reduction, loss of muscle mass, defining behavioral changes for Parkinson's disease, morphopathological changes in the brain and muscle tissue, modified circulating biomarkers, before and after moderate physical exercise.

Project objectives

1. Establishing a panel of biomarkers for inter-tissue cooperation through soluble (circulating) molecules in the in vitro "proof of concept" model, by co-cultivating human dopaminergic neurons, before and after the PB pathology induced by treatment with rotenone, and differentiated human muscle cells/sarcopenia model induced with the help of ASOs.
2. Evaluation of the effect of moderate physical exercise on the establishment and progression of muscle damage in the BP animal model by monitoring muscle and neurological parameters. 
3. Establishing a panel of circulating biomarkers in the "proof-of-concept" in vivo model, for inter-tissue cooperation through soluble molecules, as prognostic factors for the progression of BP to advanced stages and the establishment of muscle damage.
4. Establishing a panel of circulating prognostic biomarkers in BP patients with different degrees of muscle damage. 

Estimated results:

- the changes induced by Parkinson's disease on the degradation of muscle mass by determining the panel of muscle biomarkers that characterize the advanced stages of BP.

- the protective effect of physical exercise in BP, both on the changes in the brain and in the muscle tissue, on the brain-striated muscle axis. 

- characterization of the panel of modified circulating biomarkers in Parkinson's disease with associated sarcopenia: myokines, miRNAs, epigenetic and oxidative stress markers.

Project Deliverables:

Communications and scientific meetings

Bastian A., Manole E. Pathological aspects and classification criteria of inflammatory myopathies, International Pathology Conference of the “Victor Babes” Institute, Bucharest, 2-4 November 2023, Bucharest, Romania, Book of abstracts https://www.ivb.ro/wp-content/uploads/2024/01/REZUMATE-IPC_2023.pdf , Pag 53. Oral presentation

Manole E., Gaina G., Lambresu I., Ceafalan L. The involvement of Parkin in the skeletal muscle phenotype in Parkinson’s disease, International Pathology Conference of the “Victor Babes” Institute, Bucharest, 2-4 November 2023, Bucharest, Romania, Book of abstracts https://www.ivb.ro/wp-content/uploads/2024/01/REZUMATE-IPC_2023.pdf, Pag 55. Oral presentation 

Publications 

Ioghen OC, Ceafalan LC, Popescu BO. SH-SY5Y Cell Line In Vitro Models for Parkinson Disease Research-Old Practice for New Trends. J Integr Neurosci. 2023 Jan 16;22(1):20. doi: 10.31083/j.jin2201020. PMID: 36722247.

Emilia Manole, Laura C. Ceafalan, Gisela F. Gaina, Oana A. Mosoia, Mihai E. Hinescu. Myokine expression in cancer cachexia. In: Cancer research: an interdisciplinary approach. In: Rezaei, N. (eds) Cancer Research: An Interdisciplinary Approach. Interdisciplinary Cancer Research, 2023, vol 1. Springer, Cham. https://doi.org/10.1007/16833_2023_138

Ioana Lambrescu, Emilia Manole, Laura Cristina Ceafalan, Gisela Gaina. The potential benefits of drug-repositioning in muscular dystrophies. In: Potential Therapeutic strategies for muscular dystrophy. Cap. 4. Edited by Gisela Gaina, 2023, IntechOpen. ISBN: 978-1-83768-156-3, Print ISBN: 978-1-83768-155-6