SPOKE 4 - miRTOSMA - Exploring the potential of miRNAbased therapeutics to target mitochondria in spinal muscle atrophy

spoke.png

Titolo progetto: SPOKE 4 - miRTOSMA - Exploring the potential of miRNAbased therapeutics to target mitochondria in spinal muscle atrophy

Programma di finanziamento: National Center for Gene Therapy and Drugs based on RNA Technology (CN RNA & Gene Therapy). Bando a cascata CN3SP4

Responsabile scientifico: dott.ssa Camilla Bean

Ruolo del DAME: coordinator

Descrizione generale: 

  • WP 1 Recruitment of SMA patients and collection of blood samples, as well as maintenance of the generated iPSCs in the BioBank. The current number of SMA patients recruited to participate in the project is 15 patients, but the number can increase as the project progresses. This will increase the likelihood of successfully generating iPSC lines, which will also be available for future studies by being stored in the BioBank.
  • WP 1 Generation of iPSCs from peripheral blood mononuclear cells PBMCs of SMA patients.Differentiation of iPSCs in skeletal muscle cells or neuromuscular organoids. Since 15 SMA patients are enrolled in the project as blood sample donors, we will generate a corresponding number of iPSC lines.
  • WP 1 Evaluation of miRNAs expression in iPSCs differentiated into skeletal muscle cells or neuromuscular organoids. We will generate a comprehensive panel of miRNAs differentially expressed in iPSCs derived from SMA patients.
  • WP 2 Recruitment of SMA patients and collection of blood samples for iPSCs from SMA patients, as well as maintenance of the generated iPSCs in the BioBank. The current number of SMA patients recruited to participate in the project is 15 patients, but the number can increase as the project progresses. This will increase the likelihood of successfully generating iPSC lines, which will also be available for future studies by being stored in the BioBank.
  • WP 2 Cell culture maintenance and differentiation of iPSCs from SMA patients. We will develop both 2D and 3D culture system form SMA-patient derived iPSCs
  • WP 2 Treatment of iPSCs differentiated into skeletal muscle cells or neuromuscular organoids with AntagomiRs to determine if it leads to a reduction in the levels of the corresponding microRNAs. We will generate a comprehensive panel of validated miRNAs differentially expressed in iPSCs derived from SMA patients.
  • WP 2 RNA-sequencing to study the effect on global transcriptional reprogramming by antagomir. Identification of the key signaling pathway affected by SMN loss that can be targeted for RNA therapy.
  • WP 2 Identification of mitochondrial and other subcellular organelles shape changes in AntagoMirtreated iPSCs of SMA patients by transmission electron microscopy (TEM) imaging. Identification of the key mitochondrial functions affected by SMN loss that are successfully targeted by miRNA therapy.
  • WP 2 Measurement of critical bioenergetic and functional parameters of mitochondria in AntagoMirtreated iPSCs from SMA patients using the Operetta (PerkinElmer) high-content imaging system, Identification of the key mitochondrial functions affected by SMN loss that are successfully targeted by miRNA therapy.
  • WP 2 Measurement of metabolic reprogramming in AntagoMir-treated iPSCs from SMA patients using MALDI. Identification of the key mitochondrial functions affected by SMN loss that are successfully targeted by miRNA therapy.

Partner del progetto:

  • Università degli studi di Udine

Date inizio e fine progetto: 19.09.2024 – 18.09.2025

Budget totale del progetto: 245.745,00 €

Iniziativa finanziata dall’Unione europea –NextGenerationEU.