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# Mechanistic Hypotheses: Curli Amyloid Trans-BBB Seeding of α-Synuclein Aggregation

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## Hypothesis 1: LRP1-Mediated Curli Transcytosis Across the BBB

**Title:** LRP1 receptor mediates curli amyloid transcytosis

**Mechanism:** Curli fibrils (primarily CsgA monomers with conserved β-solenoid structure) bind to the low-density lipoprotein receptor-related protein 1 (LRP1) on brain microvascular endothelial cells. LRP1-mediated endocytosis packages curli into vesicles that traffic across the endothelial cytoplasm, releasing curli into the brain parenchyma via exocytosis at the abluminal membrane. Released curli directly templating α-synuclein (SNCA) monomer misfolding via cross-β-sheet complementarity.

**Key Evidence:**
- E. coli curli accelerate α-synuclein aggregation and dopaminergic neurodegeneration in M83 mice (PMID: 31101651)
- LRP1 mediates transcytosis of other amyloid species (Aβ40, α-synuclein oligomers) across BBB (PMID: 23702984)

**Testable Prediction:** siRNA knockdown of LRP1 in human iPSC-derived brain microvascular endothelial cells (BMVECs) will reduce transwell transport of purified FITC-labeled curli fibrils by >70% compared to non-targeting controls.

**Target Gene/Protein:** LRP1 (LIPR1_HUMAN)

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## Hypothesis 2: Cross-Seeding at the Neurovascular Unit via C3b Opsonization

**Title:** Complement C3 deposits on curli enabling BBB crossing

**Mechanism:** Systemic curli activates the classical complement cascade via C1q binding to curli fibrils (similar to Aβ recognition), leading to C3b opsonization. C3b-coated curli engages CR3 (CD11b/CD18) on infiltrating monocytes and neutrophils, hitchhiking across a compromised BBB. Alternatively, C3b binds CR3 on brain endothelial cells, triggering LRP1-independent transmigration. Within the brain, C3b-curli complexes directly nucleate α-synuclein fibrillization.

**Key Evidence:**
- Curli activate complement factor C3 in vitro and in vivo (PMID: 31249567)
- C3 deficiency attenuates α-synuclein pathology in MOG35-80-immunized mice (PMID: 31953283)

**Testable Prediction:** C3 knockout mice colonized with curli-producing E. coli will show reduced brain parenchymal curli (measured by RT-QuIC) and attenuated α-synuclein seeding compared to WT colonized mice.

**Target Gene/Protein:** Complement C3 (CO3_HUMAN)

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## Hypothesis 3: Bacterial Outer Membrane Vesicles Carry Curli Across the BBB

**Title:** Curli-vesicle complexes penetrate BBB intact

**Mechanism:** Curli fibers become incorporated into or

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