A Prospective Observational Study of Cerebrospinal Fluid Mitochondrial Biomarkers Measured by Flow Cytometry in Patients With Ischemic Stroke and Alzheimer Disease Controls

This prospective observational study aims to investigate cerebrospinal fluid mitochondrial biomarkers in patients with ischemic stroke and Alzheimer disease controls who undergo diagnostic lumbar puncture for clinical indications at Xuanwu Hospital, Capital Medical University.

Residual cerebrospinal fluid samples will be analyzed by flow cytometry to quantify mitochondrial content, mitochondrial membrane potential, and cellular or vesicular source-related markers. The flow cytometry panel will include MitoTracker, JC-1, and membrane-associated markers including CD45, CD41, CD24, vWF, and EAAT1.

In patients with ischemic stroke, the study will further examine whether cerebrospinal fluid mitochondrial measurements are associated with neurological severity and functional outcomes, including admission and discharge NIHSS scores and the 90-day modified Rankin Scale score. Alzheimer disease patients undergoing diagnostic lumbar puncture will serve as disease controls for biomarker comparison.

Study Overview

• Status: Recruiting
• Conditions: Ischemic Stroke; Alzheimer Disease (AD)
• Intervention / Treatment: Diagnostic test: Cerebrospinal Fluid Flow Cytometry

Detailed Description

Mitochondrial dysfunction is closely associated with ischemic brain injury, but the clinical relevance of extracellular or cell-associated mitochondrial signals in the cerebrospinal fluid of patients with ischemic stroke remains insufficiently characterized. Cerebrospinal fluid may provide a biologically informative compartment for assessing mitochondrial injury, mitochondrial membrane potential, and potential cellular sources of mitochondrial signals in neurological diseases.

This is a single-center, prospective, observational study conducted at Xuanwu Hospital, Capital Medical University. The study will enroll patients with ischemic stroke or Alzheimer disease who require diagnostic lumbar puncture as part of routine clinical evaluation between March 2026 and May 2026. No lumbar puncture will be performed solely for research purposes. After completion of clinically required testing, available residual cerebrospinal fluid will be collected for research flow cytometry analysis.

Flow cytometry will be used to evaluate cerebrospinal fluid mitochondrial content and related phenotypic features. MitoTracker will be used to detect mitochondrial signal, JC-1 will be used to assess mitochondrial membrane potential, and membrane-associated markers including CD45, CD41, CD24, vWF, and EAAT1 will be used to characterize potential leukocyte-associated, platelet-associated, neuronal/extracellular vesicle-associated, endothelial-associated, and astrocytic/glial-associated components.

The primary analysis will focus on patients with ischemic stroke and will evaluate the association between cerebrospinal fluid mitochondrial content and 90-day functional outcome measured by the modified Rankin Scale. Secondary analyses will examine associations between cerebrospinal fluid mitochondrial measurements and admission NIHSS, discharge NIHSS, and changes in NIHSS during hospitalization. Exploratory analyses will compare cerebrospinal fluid mitochondrial and membrane-marker profiles between ischemic stroke patients and Alzheimer disease controls.

• Study Type: Observational
• Enrollment (Estimated): 40

Contacts and Locations

• Study Contact: Name: Xi Chen, MD/PhD; Phone Number: 8617600104266; Email: chenxi00cx@126.com

Study Locations

• China
  • None Selected
    • Beijing, None Selected, China, 100053
      • Recruiting
      • Xuanwu Hospital
      • Contact: Xi Chen; Phone Number: +86 17600104266; Email: chenxi00cx@126.com

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

• Sampling Method: Non-Probability Sample

Study Population

The study population will include adult patients treated at Xuanwu Hospital, Capital Medical University between March 2026 and May 2026 who are diagnosed with ischemic stroke or Alzheimer disease and require diagnostic lumbar puncture for clinical indications. Only patients with available residual cerebrospinal fluid after completion of routine clinical testing will be included.

Description

Inclusion Criteria:

• Age 18 years or older. Patients treated at Xuanwu Hospital, Capital Medical University between March 2026 and May 2026.

Diagnosis of ischemic stroke or Alzheimer disease according to standard clinical diagnostic criteria.

Diagnostic lumbar puncture performed for clinical indications as part of routine medical care.

Availability of residual cerebrospinal fluid after completion of clinically required testing.

Ability to provide written informed consent, or availability of a legally authorized representative to provide consent when appropriate.

For ischemic stroke patients, availability of baseline neurological assessment and planned follow-up for 90-day modified Rankin Scale assessment.

Exclusion Criteria:

• Lumbar puncture performed solely for research purposes rather than clinical indication.

Insufficient residual cerebrospinal fluid volume for research flow cytometry analysis.

Grossly bloody or severely contaminated cerebrospinal fluid sample that precludes reliable flow cytometry analysis.

Known central nervous system infection, malignant meningitis, or other inflammatory or neoplastic condition that, in the investigator's judgment, may substantially confound cerebrospinal fluid mitochondrial measurements.

Inability to obtain informed consent from the participant or legally authorized representative.

Missing key clinical outcome data, including admission NIHSS, discharge NIHSS, or planned 90-day mRS follow-up for ischemic stroke participants.

Any condition judged by the investigator to make the participant unsuitable for inclusion in the study.

Study Plan

How is the study designed?

Number of groups / cohorts

2

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Ischemic Stroke Cohort; Patients diagnosed with ischemic stroke who undergo diagnostic lumbar puncture for clinical indications at Xuanwu Hospital, Capital Medical University. Residual cerebrospinal fluid will be analyzed by flow cytometry to assess mitochondrial content, mitochondrial membrane potential, and membrane-associated markers. Clinical outcomes will include admission NIHSS, discharge NIHSS, and 90-day modified Rankin Scale score.	Diagnostic test: Cerebrospinal Fluid Flow Cytometry; Residual cerebrospinal fluid obtained during clinically indicated diagnostic lumbar puncture will be analyzed by flow cytometry. The assay will measure mitochondrial signal using MitoTracker, mitochondrial membrane potential using JC-1, and membrane-associated markers including CD45, CD41, CD24, vWF, and EAAT1. The study does not assign participants to any treatment or diagnostic procedure beyond routine clinical care.
Alzheimer Disease Control Cohort; Patients diagnosed with Alzheimer disease who undergo diagnostic lumbar puncture for clinical indications at Xuanwu Hospital, Capital Medical University. Residual cerebrospinal fluid will be analyzed by the same flow cytometry panel and used as a disease control group for comparison of cerebrospinal fluid mitochondrial and membrane-marker profiles.	Diagnostic test: Cerebrospinal Fluid Flow Cytometry; Residual cerebrospinal fluid obtained during clinically indicated diagnostic lumbar puncture will be analyzed by flow cytometry. The assay will measure mitochondrial signal using MitoTracker, mitochondrial membrane potential using JC-1, and membrane-associated markers including CD45, CD41, CD24, vWF, and EAAT1. The study does not assign participants to any treatment or diagnostic procedure beyond routine clinical care.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Association Between Cerebrospinal Fluid Mitochondrial Content and 90-Day Modified Rankin Scale Score in Ischemic Stroke	The primary outcome is the association between cerebrospinal fluid mitochondrial content, measured by flow cytometry using MitoTracker-positive events or MitoTracker fluorescence intensity, and functional outcome at 90 days assessed by the modified Rankin Scale. The modified Rankin Scale ranges from 0 to 6, with higher scores indicating greater disability or death.	From baseline cerebrospinal fluid collection to 90 days after stroke onset or hospital admission

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Association Between Cerebrospinal Fluid Mitochondrial Content and Admission NIHSS Score	The association between cerebrospinal fluid mitochondrial content and baseline neurological deficit severity will be assessed using the National Institutes of Health Stroke Scale. Higher NIHSS scores indicate more severe neurological impairment.	At hospital admission or baseline clinical assessment
Association Between Cerebrospinal Fluid Mitochondrial Content and Discharge NIHSS Score	The association between cerebrospinal fluid mitochondrial content and neurological deficit at discharge will be assessed using the NIHSS score recorded at hospital discharge.	At hospital discharge, up to 7 days after admission
Cerebrospinal Fluid Mitochondrial Membrane Potential Measured by JC-1	Mitochondrial membrane potential in cerebrospinal fluid-associated mitochondrial events will be assessed by JC-1 flow cytometry. JC-1 fluorescence parameters will be used to characterize mitochondrial polarization status.	At baseline cerebrospinal fluid collection
Comparison of Cerebrospinal Fluid Mitochondrial Biomarkers Between Ischemic Stroke and Alzheimer Disease Controls	Cerebrospinal fluid mitochondrial content, JC-1-related mitochondrial membrane potential measures, and membrane-associated marker profiles will be compared between ischemic stroke patients and Alzheimer disease controls.	At baseline cerebrospinal fluid collection

Collaborators and Investigators

• Sponsor: Capital Medical University

Study record dates

Study Major Dates

• Study Start (Actual): March 1, 2026
• Primary Completion (Estimated): August 31, 2026
• Study Completion (Estimated): November 30, 2026

Study Registration Dates

• First Submitted: May 15, 2026
• First Submitted That Met QC Criteria: May 15, 2026
• First Posted (Actual): May 22, 2026

Study Record Updates

• Last Update Posted (Actual): May 22, 2026
• Last Update Submitted That Met QC Criteria: May 15, 2026
• Last Verified: May 1, 2026

More Information

Terms related to this study

• Keywords: Mitochondria; Cerebrospinal fluid; NIHSS; MitoTracker; Stroke outcome
• Additional Relevant MeSH Terms: Cerebrovascular Disorders; Brain Diseases; Central Nervous System Diseases; Nervous System Diseases; Vascular Diseases; Cardiovascular Diseases; Mental Disorders; Neurocognitive Disorders; Dementia; Tauopathies; Neurodegenerative Diseases; Stroke; Ischemic Stroke; Alzheimer Disease
• Other Study ID Numbers: CSF-MITO

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO
• IPD Plan Description: The study is proceeding.

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No