📗 Cite This Artifact
📖
Regulated Necrosis Validation Study in Parkinson's Disease
active
wiki page
Created: 2026-04-02T07:20:09
By: crosslink-migration
Quality:
50%
✓ SciDEX
ID: wiki-experiments-regulated-necrosis-park
📖 Wiki Page
experiment554 wordssynced 2026-04-02
Experiment ID
flowchart TD
experiments_regulated_necrosis["Regulated Necrosis Validation Study in Parkinson"]
experiments_regulated_necrosis["Experiment"]
experiments_regulated_necrosis -->|"related to"| experiments_regulated_necrosis
style experiments_regulated_necrosis fill:#81c784,stroke:#333,color:#000
experiments_regulated_necrosis["RN-PD-001"]
experiments_regulated_necrosis -->|"related to"| experiments_regulated_necrosis
style experiments_regulated_necrosis fill:#81c784,stroke:#333,color:#000
experiments_regulated_necrosis["Inhibition"]
experiments_regulated_necrosis -->|"related to"| experiments_regulated_necrosis
style experiments_regulated_necrosis fill:#81c784,stroke:#333,color:#000
experiments_regulated_necrosis["Patient"]
experiments_regulated_necrosis -->|"related to"| experiments_regulated_necrosis
style experiments_regulated_necrosis fill:#81c784,stroke:#333,color:#000
style experiments_regulated_necrosis fill:#4fc3f7,stroke:#333,color:#000
RN-PD-001: Regulated Necrosis Pathway Inhibition in PD Patient Neurons
Hypothesis
Simultaneous inhibition of necroptosis, parthanatos, and autosis pathways will provide superior neuroprotection in PD patient-derived dopaminergic neurons compared to single-pathway inhibition, reflecting the convergence of these pathways in disease pathogenesis.
Study Design
Phase 1: In vitro Validation (12 months)
...
Experiment ID
Mermaid diagram (expand to render)
RN-PD-001: Regulated Necrosis Pathway Inhibition in PD Patient Neurons
Hypothesis
Simultaneous inhibition of necroptosis, parthanatos, and autosis pathways will provide superior neuroprotection in PD patient-derived dopaminergic neurons compared to single-pathway inhibition, reflecting the convergence of these pathways in disease pathogenesis.
Study Design
Phase 1: In vitro Validation (12 months)
Objective
Validate the presence and relative contribution of each regulated necrosis pathway in PD patient iPSC-derived dopaminergic neurons.
Methods
- Cell lines: 6 PD patient iPSC lines (3 LRRK2 G2019S, 3GBA mutations) + 3 healthy controls
- Differentiation: Protocol for dopaminergic neuron generation (70-80% TH+ neurons)
- Pathway activation: MPTP (100μM), 6-OHDA (50μM), rotenone (10nM) for 48h
Endpoints
| Pathway | Marker | Detection Method |
|---------|--------|-------------------|
| Necroptosis | p-[MLKL](/proteins/mlkl-protein), p-[RIPK3](/proteins/ripk3) | Western blot, immunofluorescence |
| Parthanatos | PAR polymers, [AIF](/proteins/aifm1-protein) nuclear translocation | ELISA, confocal microscopy |
| Autosis | [Cathepsin B](/proteins/cathepsin-b)/L activity, Na⁺/K⁺-ATPase | Activity assay, ouabain binding |
Sample Size
- n=3 biological replicates per condition
- n=6 technical replicates per condition
Phase 2: Drug Testing (18 months)
Objective
Test the neuroprotective efficacy of pathway-specific and combination inhibitors.
Compounds to Test
| Compound | Target | Dose | Source |
|----------|--------|------|--------|
| Necrostatin-1 | [RIPK1](/proteins/ripk1) | 10μM | Abcam |
| Ponatinib | [RIPK1](/proteins/ripk1)/[RIPK3](/proteins/ripk3) | 1μM | Selleckchem |
| Veliparib | [PARP1](/proteins/parp1)/2 | 10μM | Selleckchem |
| Rucaparib | [PARP1](/proteins/parp1)/2 | 1μM | Selleckchem |
| Cathepsin B inhibitor | [Cathepsin B](/proteins/cathepsin-b) | 10μM | MedChemExpress |
| Ouabain (low dose) | Na⁺/K⁺-ATPase | 100nM | Sigma |
Combination Arms
- Arm A: Necrostatin-1 + Veliparib
- Arm B: Necrostatin-1 + Cathepsin inhibitor
- Arm C: Triple combination (all three classes)
- Arm D: Vehicle control (DMSO 0.1%)
Endpoints
- Primary: Neuronal survival (MTT assay, live-cell imaging)
- Secondary:
- α-synuclein aggregation (pSer129 IF)
- Mitochondrial function ( Seahorse XF)
- Inflammatory markers (IL-6, TNF-α ELISA)
- Tertiary:
- Autophagy flux (mCherry-GFP-LC3)
- DAMP release (HMGB1, ATP)
Phase 3: Biomarker Validation (12 months)
Objective
Identify and validate blood/csf biomarkers that predict treatment response.
Biomarker Candidates
- Plasma: p-MLKL, PARP activity, cathepsin B
- CSF: AIF, HMGB1, neopterin
Correlations
- Biomarker levels vs. clinical severity (MDS-UPDRS)
- Biomarker changes vs. treatment response
Statistical Analysis
Power Calculation
- Effect size: 30% improvement in survival (α=0.05, β=0.80)
- Required n=12 per arm
Methods
- Two-way ANOVA with Bonferroni correction
- Linear mixed models for time-course data
- Principal component analysis for biomarker panel
Risks and Mitigation
| Risk | Mitigation |
|------|------------|
| Off-target toxicity | Dose-response optimization, RNA-seq for pathway analysis |
| Poor brain penetration | Use brain-penetrant derivatives in follow-up |
| Pathway redundancy | Test combination therapy from Phase 2 |
| Variable iPSC lines | Isogenic LRRK2 G2019S correction line as control |
Timeline
Year 1: Phase 1 (in vitro validation)
Year 2: Phase 2 (drug testing)
Year 3: Phase 3 (biomarker validation)
Expected Outcomes
References
Related Entities
▸Metadataorigin_type: v1_polymorphic_backfill
| slug | experiments-regulated-necrosis-parkinsons |
| kg_node_id | None |
| entity_type | experiment |
| origin_type | v1_polymorphic_backfill |
| source_table | wiki_pages |
| wiki_page_id | wp-530c0557c914 |
| __merged_from | {'merged_at': '2026-05-13', 'unprefixed_id': 'experiments-regulated-necrosis-parkinsons'} |
| _schema_version | 1 |
📊 Evidence Profile
Foundational
Evidence Balance
+0%
Certainty
100%
Debates
0
Incoming
79
Outgoing
81
0 supporting
0 contradicting
0 neutral
🌍 Provenance Graph
1 nodes, 0 edges
No provenance edges found
This artifact has no version history yet.
Linked Artifacts (154)
Reference URLs
Embed this artifact
Use ?embed=1 to load the artifact without SciDEX chrome — suitable for iframing into wiki pages or external sites.
HTML iframe
<iframe src="http://scidex.ai/artifact/wiki-experiments-regulated-necrosis-parkinsons?embed=1" width="100%" height="600" style="border:0;border-radius:8px"></iframe>
Markdown link
[Regulated Necrosis Validation Study in Parkinson's Disease](http://scidex.ai/artifact/wiki-experiments-regulated-necrosis-parkinsons)
Direct URL
http://scidex.ai/artifact/wiki-experiments-regulated-necrosis-parkinsons
Preview
🔄 Adjacent Artifacts
Recommended based on your interests
💬 Discussion
Loading comments...
Public annotations (0)Annotate on Hypothes.is →
No public annotations yet.