Research at Biopathio
Our research program focuses on solving the fundamental formulation and targeting problems that limit CNS gene therapy delivery. We publish findings as preprints and conference presentations as work matures.
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Four Focus Areas
Each research area addresses a distinct technical barrier in CNS gene therapy delivery.
LNP Formulation for CNS Delivery
Ionizable lipid pKa screening (5.8–6.2 window) against neural cell endosomal escape assays, helper lipid molar ratio optimization, and characterization protocols for CNS applications including DLS, cryo-TEM, RiboGreen EE, and Gal8-based endosomal escape quantification. Direct comparison of formulation performance in hepatocyte vs. primary cortical neuron models.
Blood-Brain Barrier Transcytosis
Mechanistic studies of receptor-mediated transcytosis at the BBB. TfR1 and LRP1 pathway characterization in hCMEC/D3 transwell models with TEER validation. Targeting ligand density optimization, dual-receptor strategies, and Papp quantification. In vitro transcytosis data for TfR1-targeted, LRP1-targeted, and dual-targeting formulations.
CRISPR Nucleic Acid Encapsulation
Encapsulation efficiency and cargo integrity characterization for Cas9 RNP, mRNA + sgRNA dual-cargo, ABE8e mRNA + sgRNA, CBE4max mRNA + sgRNA, and PE2 + pegRNA formulations. N/P ratio optimization (4–6 target range for dual-cargo), nucleic acid:lipid mass ratio titration, and release kinetics in neural cell endosomal pH conditions.
In Vitro CNS Cell Models
Development and validation of in vitro models for evaluating CNS-targeted LNP delivery: SH-SY5Y differentiated neurons (endosomal escape and editing efficiency screen), primary rat cortical neurons DIV7–10 (functional editing readout), hCMEC/D3 transwell BBB models (transcytosis efficiency), and neuron-astrocyte co-culture systems for cell-type-selective uptake assessment.
CNS Rare Disease Targets
Biopathio's platform development is informed by three CNS rare disease contexts where LNP delivery is most needed and the genetic targets are best characterized.
Huntington's Disease (HTT)
CAG trinucleotide repeat expansion in HTT exon 1 causes polyglutamine expansion and progressive striatal and cortical neurodegeneration. CRISPR silencing of mutant HTT allele via repeat-adjacent guide RNAs has demonstrated in vitro knockdown in patient-derived iPSC neurons. The delivery challenge: striatal medium spiny neurons require systemic delivery across the BBB, which no non-viral platform has demonstrated at therapeutic doses in primates.
Friedreich's Ataxia (FXN)
GAA trinucleotide repeat expansion in FXN intron 1 causes epigenetic silencing of frataxin, a mitochondrial iron-sulfur cluster protein. CRISPRa and dCas9-TET-based epigenetic reactivation of FXN has restored frataxin expression to near-normal levels in patient-derived FA neurons in vitro. The delivery challenge: primary neurodegeneration occurs in dorsal root ganglia and spinocerebellar pathways — anatomical targets requiring both CNS and peripheral nervous system access.
CLN3 Batten Disease
A 1.02 kb deletion spanning CLN3 exons 7–8 accounts for ~85% of disease alleles. CRISPR exon-skipping strategies producing an in-frame partial CLN3 protein have shown efficacy in murine CLN3 models. The delivery challenge: CLN3 neurodegeneration is diffuse — involving cortical, subcortical, and cerebellar neurons — requiring broad parenchymal distribution after BBB crossing rather than targeted regional delivery.
Early-Stage Scientific Output
Biopathio's research is published as preprints and conference presentations. As a 2024-founded preclinical company, our publication record reflects our formulation and mechanism work to date.
View All PublicationsInterested in collaborating on CNS delivery research?
We are open to academic and industry research collaborations in LNP CNS delivery.