T cell responses at diagnosis of amyotrophic lateral sclerosis predict disease progression
Documentation files
Documentation files
Citation and access
Citation and access
Data access level:
Creator/Principal investigator(s):
Research principal:
Data contains personal data:
Yes
Type of personal data:
human single cell RNA sequencing data
Code key exists:
Yes
Sensitive personal data:
Yes
Citation:
Language:
Method and outcome
Method and outcome
Unit of analysis:
Population:
Participants were recruited from the bigger Stockholm area as part of the case-control studies ALSrisc and StopMS. For more Information on ALSrisc please visit the website: https://ki.se/en/imm/amyotrophic-lateral-sclerosis-als. For more information on StopMS please visit the website: https://ki.se/en/cns/fredrik-piehls-research-group
Study design:
- Observational study
- Case-control study
Description of study design:
Human material: Immune cells isolated from cerebral spinal fluid (CSF). CSF samples were collected from five patients with newly diagnosed ALS, and four controls (two patients with normal pressure hydrocephalus, one patient with cervical radiculopathy, and one healthy control). Singe cell sequencing platform: 5’ scRNA-seq + V(D)J TCR repertoire sequencing using the 10x Genomics platform. For more general inofrmation on the 10x Genomic platform please see here: https://www.10xgenomics.com/support/single-cell-immune-profiling User guids for 5’ scRNA-seq and V(D)J TCR repertoire can be found here respectively: https://assets.ctfassets.net/an68im79xiti/1ihALQkNrgD83PmBYZ4SrW/e94dbc40c2a6c64940ebc82eca9e9334/CG000507_ChromiumSingleCell5_v2_FeatureBarcode_Automation_UG_Rev_A.pdf https://assets.ctfassets.net/an68im79xiti/2JzjFJTfslSYe6GF9eh5TL/c228c4cc6b84c4f17fc6c25c8d7a5448/CG000207_ChromiumNextGEMSingleCellV_D_J_ReagentKits_v1.1_UG_Rev_G.pdf
Description of sampling:
CSF samples were collected at the time of diagnosis from ALS patients and controls at Karolinska University Hospital. 16mL CSF sample was collected in two 10mL plastic collection tubes (Sarstedt) by lumbar puncture. Within a 3-hour time window samples were transported at 4°C for processing. 1. CSF sample was centrifuged at 300g, 10min, 4°C 2. Supernatant was removed expect for around 500ul of CSF (here tubes with a visible blood contammination were droped from the analysis) 3. When more than one collection tube was used, sample were combined in one collection tube. 4. CSF samples were washed with cold PBS + 0.5% BSA (w/o ETDA) by filling the collection tube up to 10 ml. 5. CSF samples was centrifuge at 300g, 10min, 4°C. 6. Supernatant was removed, cell resuspended in 500ul of cold PBS + 0.5% BSA, and transfer sample to a low binding RNA tube 7. Samples were centrifuge at 300g, 10min, 4°C. 8. supernatant was removed and cells were re-suspended cell in the remaining supernatant (~50µl) For the remaining steps, we followed the manufacture's protocol mentioned above.
Time period(s) investigated:
Number of individuals/objects:
9
Description of the response rate/participation rate:
Sample were collected from 9 participants as a one time measurement.
Geographic coverage
Geographic coverage
Geographic location:
Administrative information
Administrative information
Responsible department/unit:
Institute of Environmental Medicine [C6]
Ethics Review:
Swedish Ethical Review Authority - DNRs 2014/1815-31/4, 2018-1065/31 and DNRs 2009/2107-31/2 and 2021-02060
Funding
Funding
Funding agency:
- Swedish Research Council
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Award number:
2019-01088_VR
Award title:
Unravelling the Interplay between Metabolism, Gut Microbiome and Adaptive Immunity in Amyotrophic Lateral Sclerosis
Funding information:
Amyotrophic lateral sclerosis (ALS) is devastating neurodegenerative disorder that in lack of effective treatments leads to death within a few years of diagnosis. ALS is increasingly recognized as a systemic disease affecting not only the nervous system but also other physiological aspects. We hypothesize that there is a disease-specific interplay between metabolism, gut microbiome and adaptive immunity that substantially contributes to the etiology of ALS. Our aim is therefore to explore such interplay, and to assess the effectiveness of a treatment regimen that specifically targets it. Using a population-based case-control study, we will first characterize the complex interactions between metabolism, microbiome, and immunity in ALS, through comprehensive proteomic, metagenomic and immune-response profiling. We will then use established ALS disease models to understand the usefulness of combining a high-caloric diet with a fecal microbiota transplant in disease prevention and treatment. The proposed research is unique as it 1) combines innovative molecular platforms with a high-quality epidemiological study design, 2) uses a novel strategy of investigating multiple aspects of human physiology, and 3) offers a possibility to directly translate findings between human observational and animal experimental studies. The ultimate goal is to advance our knowledge about ALS as a disease, and to identify novel and highly warranted preventive and therapeutic targets for it.
