Effect of Physical Therapy on NLRP3 Inflammasome Activation and Muscle Atrophy in Critical Illness Myopathy (PT-NLRP3-CIM).
试验速览
- 阶段
- 不适用
- 状态
- 招募中
- 发起方
- 入组人数
- 24
- 试验地点
- 6
- 主要终点
- Relative mRNA expression of NLRP3, IL-1β, and IL-18 in muscle tissue (RT-qPCR)
研究概览
简要总结
The goal of this clinical trial is to study whether physical therapy can reduce NLRP3 inflammasome activation and muscle atrophy in patients with critical illness myopathy (CIM). It will also explore the role of NLRP3 inflammasome in the pathophysiology of CIM.
The main questions this study aims to answer are:
Is NLRP3 inflammasome activation associated with muscle atrophy through the upregulation of atrogenes?
Does physical therapy attenuate NLRP3 inflammasome activation in skeletal muscle, thereby contributing to the prevention or reduction of muscle atrophy in CIM?
Researchers will compare enhanced physical therapy using servo-assisted bed cycling (Motomed Letto®) in critically ill patients at risk of developing CIM during the early phase of ICU stay to conventional physical therapy (standard physiotherapy), to assess whether physical therapy reduces NLRP3 inflammasome activation and muscle degradation.
Participants will:
Be randomized to receive either conventional physical therapy or enhanced physical therapy (Motomed Letto®) for 7 consecutive days. A control group of patients without CIM will also be included.
Undergo assessments of NLRP3 activity, muscle atrophy markers, and transcriptomic profiles from serum and vastus lateralis muscle biopsies.
Be clinically evaluated using the SOFA scale and muscle ultrasound for CIM diagnosis.
Be followed up for changes in muscle strength and physical functionality.
Provide sociodemographic and clinical information to be recorded throughout the study.
详细描述
Critical illness myopathy (CIM) is a frequent complication in patients admitted to intensive care units (ICUs), characterized by symmetric proximal muscle weakness and respiratory muscle involvement. It has been linked to increased mortality, prolonged hospital stays, and long-term physical disability. CIM may also contribute to post-intensive care syndrome (PICS), which includes persistent cognitive, psychological, and physical impairments. In Chile, approximately 40% of patients with critical illness due to COVID-19 developed this syndrome.
Although several molecular mechanisms have been proposed, the precise etiopathogenesis of CIM remains unclear. Muscle atrophy and contractile dysfunction are hallmark features of CIM. The ubiquitin-proteasome system (UPS) and the upregulation of atrogenes such as MuRF1 and Atrogin-1 have been implicated in its development. In murine models of denervation and sepsis, the NLRP3 inflammasome-a multiprotein complex involved in innate immunity and IL-1β/IL-18 secretion-has been shown to promote muscle atrophy via activation of atrophy-related genes.
Evidence suggests that physical therapy can modulate inflammation at the skeletal muscle level, including downregulation of NLRP3 inflammasome components and IL-1β expression. Mechanical silencing, a major modifiable risk factor in CIM, can be mitigated by early mobilization strategies. However, it remains unknown whether physical therapy directly reduces NLRP3 inflammasome activation and associated muscle atrophy in patients with CIM.
This clinical trial is designed to test the hypothesis that physical therapy decreases NLRP3 inflammasome activity and reduces skeletal muscle atrophy in critically ill patients with CIM. In the early stage of ICU admission, sixteen patients at risk of CIM will be randomized to receive either conventional physical therapy or an enhanced protocol that includes servo-assisted motorized movement therapy (Motomed Letto®), twice daily for 60 minutes. In addition, eight patients without CIM will serve as non-CIM controls.
Muscle biopsy samples from the vastus lateralis will be analyzed to assess histological evidence of muscle atrophy, structural alterations in cellular organelles, and expression of atrophy-related genes. Quantitative real-time PCR (RT-qPCR) will be used to measure mRNA levels of atrogenes, and Western blot will assess protein expression of key mediators of NLRP3 inflammasome signaling. Transcriptomic analysis will be conducted using microarray profiling.
研究设计
- 研究类型
- Interventional
- 分配方式
- Randomized
- 干预模型
- Parallel
- 主要目的
- Basic Science
- 盲法
- None
入排标准
- 年龄范围
- 18 Years 至 —（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •Medical diagnosis of sepsis upon ICU admission.
- •Receiving invasive mechanical ventilation with a projected requirement ≥7 days.
- •SOFA score ≥8 for three consecutive days within the first five days of ICU admission.
排除标准
- •Neurocritical illness.
- •Prior malnutrition or cachexia.
- •Pre-existing neuromuscular disease.
- •Coagulopathy (severe liver disease or continuous dialysis).
- •Thrombocytopenia <20,000 platelets/μL.
- •Prior Clinical Frailty Scale ≥
- •Lower limb amputation or fractures.
- •Ongoing chemotherapy.
- •Pregnancy.
- •Uncontrolled epilepsy.
- •Allergy to ultrasound gel.
- •Prior prolonged corticosteroid therapy.
- •Expected ICU stay <7 days.
- •Imminent death.
- •Legal guardian refusal to provide informed consent.
研究组 & 干预措施
Enhanced or Additional Physical Therapy Group
Critically ill patients at risk of developing CIM will receive enhanced (additional) physical therapy consisting of standard physiotherapy plus servo-assisted lower-limb cycling using the Motomed Letto® device. Therapy will be delivered twice daily, 60 minutes per session, for 7 consecutive days during the early ICU stay. Muscle biopsies from the vastus lateralis will be collected for histological and molecular analyses.
干预措施: Motomed Letto® servo-assisted cycling + standard physiotherapy (Device)
Conventional Physical Therapy Group
Critically ill patients at risk of CIM will receive standard physiotherapy delivered according to institutional ICU rehabilitation protocols. The intervention is delivered at the same frequency and clinical stage as the enhanced therapy group.
干预措施: Standard physiotherapy (Behavioral)
结局指标
主要结局
Relative mRNA expression of NLRP3, IL-1β, and IL-18 in muscle tissue (RT-qPCR)
时间窗: Baseline and Day 7 (on the final day of intervention)
mRNA levels of NLRP3, IL-1β, and IL-18 will be quantified using RT-qPCR from muscle biopsy samples.
Protein expression of phospho-p65S536 and total p65 (Western blot)
时间窗: Baseline and Day 7 (on the final day of intervention)
Protein levels of phospho-p65 Serine 536 and total p65 will be assessed by Western blot to evaluate NF-κB pathway activation.
Ratio of cleaved/uncleaved caspase-1, IL-1β, and GSDMD-NT (Western blot)
时间窗: Baseline and Day 7 (on the final day of intervention)
Cleaved forms of IL-1β, caspase-1, and GSDMD-NT will be quantified by Western blot and normalized to precursor forms.
Plasma concentrations of IL-1β and IL-18 (ELISA)
时间窗: Baseline and Day 7 (on the final day of intervention)
Plasma levels of IL-1β and IL-18 will be measured using ELISA.
Expression of oxidative stress markers and cathepsin B (Western blot)
时间窗: Baseline and Day 7 (on the final day of intervention)
Protein carbonylation, nitration, and cathepsin B expression will be analyzed by Western blot.
Expression of cathepsin B (RT-qPCR)
时间窗: Baseline and Day 7 (on the final day of intervention)
Cathepsin B expression will be analyzed by RT-qPCR.
Ultrastructural mitochondrial damage and lysosomal vacuolization (TEM)
时间窗: Baseline and Day 7 (on the final day of intervention)
Mitochondrial damage and lysosomal vacuole volume will be assessed by transmission electron microscopy.
Muscle fiber diameter (immunofluorescence)
时间窗: Baseline and Day 7 (on the final day of intervention)
Fiber diameter in 10 µm cryosections will be measured using anti-laminin immunofluorescence.
mRNA expression of atrogenes: MuRF1, Atrogin-1, MUSA1, TRIM62, TRIM32
时间窗: Baseline and Day 7 (on the final day of intervention)
Atrogene mRNA levels will be quantified using RT-qPCR from muscle biopsies.
Myosin/actin ratio (Western blot)
时间窗: Baseline and Day 7 (on the final day of intervention)
Myosin and actin protein levels will be analyzed by Western blot to assess the molecular diagnosis of CIM.
Sarcomeric organization (TEM)
时间窗: Baseline and Day 7 (on the final day of intervention)
Evaluation of sarcomeric structure in cryosections using transmission electron microscopy.
次要结局
- Differential expression of genes related to atrophy and inflammasome activation (microarray or nanopore-based RNA sequencing )(Baseline (prior to intervention) and Day 7 of intervention)
- Correlation between molecular findings and CIM clinical diagnosis(From recovery of consciousness (in ICU or follow-up) through 90-day follow-up)
研究者
Oscar Arellano
Principal Investigator, PhD Candidate in Biomedical Sciences, University of Chile
University of Chile