Neuroimmunology

Neuroimmunology & Neuroinflammation

Investigating the intersection of innate immunity and neurodegeneration in human iPSC-derived models. My neuroimmunology program dissects how pro-inflammatory cytokines (IFN-γ, TNF-α, IL-1β) drive neuronal vulnerability, α-synuclein aggregation, and lysosomal dysfunction — and how microglia and astrocytes modulate these pathological cascades. This work has yielded first-author publications in Nature Neuroscience and Cell Reports, with direct translational applications for anti-inflammatory neuroprotection strategies.

Cytokines

Cytokine-Driven Neuronal Vulnerability

Systematic investigation of how pro-inflammatory cytokines impact iPSC-derived dopaminergic neuron health. IFN-γ exposure induces MHC-I upregulation, antigen presentation machinery activation, and downstream lysosomal dysfunction. TNF-α and IL-1β act synergistically to promote α-synuclein phosphorylation and Lewy body-like inclusion formation. These findings establish a mechanistic framework connecting peripheral inflammation to neuronal pathology in Parkinson's disease.

Key Findings

Cytokine Pathways Characterized (IFN-γ, TNF-α, IL-1β)

4.2×

α-Syn Inclusion Increase (IFN-γ + PFF)

72h

Optimal Treatment Window

>85%

TH+ Neuron Purity

Cytokine Impact on iPSC-DA Neurons

Quantitative Data

Cytokine-induced pathology across multiple readouts

Data from n=3–4 biological replicates. α-Syn inclusions quantified by immunocytochemistry (cells with ≥2 inclusions). Viability by CellTiter-Glo luminescence. MHC-I expression by flow cytometry (fold-change MFI vs vehicle).

Image Gallery

IFN-γ-treated iPSC-DA neurons — IFN-γ–treated iPSC-DA neurons showing α-synuclein inclusion formation (green) with nuclear DAPI (blue)

IFN-γ-induced autophagy dysregulation — IFN-γ–induced autophagy dysregulation: LC3B puncta accumulation in dopaminergic neurons

Electron microscopy of lysosomal dysfunction — Electron microscopy revealing IFN-γ–induced lysosomal morphological dysfunction

Co-Culture

Neuron–Glia Co-Culture Systems

Developed tri-culture systems combining iPSC-derived dopaminergic neurons with microglia and astrocytes to model the neuroimmune microenvironment. Microglia amplify α-synuclein inclusion formation through pro-inflammatory cytokine secretion and direct phagocytic interactions. Astrocytes provide neuroprotection via glutathione transfer and metabolic support, but become neurotoxic when activated by microglial TNF-α. These co-culture platforms enable screening of anti-inflammatory therapeutics in physiologically relevant contexts.

Key Findings

Cell Types in Tri-Culture (Neurons + Microglia + Astrocytes)

2.8×

Microglia-Enhanced Inclusion Formation

45%

Astrocyte-Mediated Neuroprotection

15+

Cytokines Profiled by MSD

Co-Culture Effects on Neuronal Pathology

Quantitative Data

Multi-cellular effects across readouts in tri-culture

n=3 independent differentiations. α-Syn inclusions: % cells with ≥2 punctate deposits. Cell Death: % PI+ or annexin V+. Cytokine Release: pg/mL in culture supernatant (48h).

Image Gallery

iPSC-derived microglia in co-culture — iPSC-derived microglia (IBA1+, magenta) interacting with dopaminergic neurons (TH+, green) in co-culture

Astrocyte co-culture with dopaminergic neurons — Astrocyte co-culture (GFAP+, red) with dopaminergic neurons showing neuroprotective phenotype

Receptor quenching assay for microglial signaling — Receptor quenching assay: microglial cytokine receptor engagement and downstream signaling

Receptors

Neuroimmune Receptor Dissection

Systematic characterization of cytokine receptor expression and signaling in iPSC-derived dopaminergic neurons. Identified IFNGR1, TNFR2, and IL1R1 as the dominant receptor subtypes mediating inflammatory neurodegeneration. Confocal imaging confirmed surface receptor localization and internalization dynamics upon ligand binding. These receptor maps guided the selection of anti-cytokine biologics (anti-TNF-α, anti-IL-1β, anti-IL-6) that demonstrated dose-dependent neuroprotection.

Key Findings

Key Receptors Characterized (IFNGR1, TNFR2, IL1R1)

IFNGR1

Dominant Receptor in DA Neurons

60%

Neuroprotection with Anti-TNF Biologics

Biologics Screened

Cytokine Receptor Expression in iPSC-DA Neurons

Quantitative Data

Mean fluorescence intensity (MFI) by flow cytometry

Flow cytometry quantification of surface-level receptor expression. n=3 donors, 10,000+ live, single cells per sample. Isotype control background subtracted.

Image Gallery

IFNGR1 surface expression (63×) ↗ expand

Lysosomes

Inflammation-Induced Lysosomal Dysfunction

IFN-γ signaling triggers progressive lysosomal dysfunction in dopaminergic neurons, characterized by LAMP1+ compartment enlargement, cathepsin activity reduction, and impaired autophagic flux. This creates a feed-forward loop: dysfunctional lysosomes fail to clear α-synuclein aggregates, which further compromise lysosomal membrane integrity (detected by galectin-3 puncta). Electron microscopy revealed multilamellar body accumulation and lysosomal membrane rupture in IFN-γ–treated neurons.

Key Findings

3.5×

LAMP1+ Compartment Enlargement

55%

Cathepsin D Activity Reduction

2.8×

Galectin-3 Puncta Increase

48h

Onset of Lysosomal Pathology

Lysosomal Dysfunction Timeline (IFN-γ Treatment)

Quantitative Data

Time-dependent changes in lysosomal markers

Kinetic measurements from confocal microscopy. LAMP1 Intensity: fold-change in mean punctal fluorescence. Cathepsin Activity: substrate cleavage assay (luminescence). Galectin-3 Puncta: quantified per cell by automated image analysis. n=30–50 neurons per timepoint.

Image Gallery

Lysosomal swelling in PFF-treated neurons: LAMP1 (green), WGA membrane (red)

Galectin-3 puncta indicating lysosomal disruption — Galectin-3 puncta indicating lysosomal membrane permeabilization in IFN-γ–treated neurons

Electron microscopy of multilamellar bodies — Electron microscopy: multilamellar body accumulation and lysosomal membrane disruption

Biomarkers

Neuroimmune Biomarker Profiling

Comprehensive MSD multiplex profiling of secreted cytokines, chemokines, and neurodegeneration markers across inflammatory treatment paradigms. Established pharmacodynamic biomarker panels for monitoring anti-inflammatory therapeutic efficacy. Key readouts include phospho-α-synuclein (pS129), NfL, GFAP, and a 15-plex cytokine panel enabling simultaneous quantification of inflammatory and neurotoxic cascades.

Key Findings

MSD Multiplex Analytes

pS129

Key PD Biomarker

Sample Types (Media, Lysate, CSF-analog)

R²>0.99

Standard Curve Quality

Neuroimmune Biomarker Panel — Fold Change (IFN-γ vs Vehicle)

Quantitative Data

Comprehensive multiplex biomarker response to IFN-γ

MSD V-PLEX platform. n=4 biological replicates. Vehicle baseline = 1.0 (center). IFN-γ treatment (24h, 10 ng/mL). Axes represent fold-change relative to unstimulated control.

Image Gallery

Phospho-α-synuclein accumulation — Phospho-α-synuclein (pS129) accumulation time-course following dual PFF + IFN-γ treatment

Western blot biomarker panel — Western blot panel: inflammatory signaling markers across treatment conditions

Neuroimmunology Experimental Pipeline

STEP 01

iPSC Differentiation

DA neurons, microglia, astrocytes (30–60 days)

STEP 02

Inflammatory Challenge

Cytokine treatments, PFF seeding, co-culture

STEP 03

Multi-Modal Readout

ICC, flow, MSD, Western, calcium imaging

STEP 04

Pathway Dissection

Receptor mapping, signaling cascades, gene expression

STEP 05

Therapeutic Screen

Anti-inflammatory biologics, small molecules, neuroprotection