Layman Explanation
Müller glia help the retina. We use short‑lived mRNA (no genome changes) to switch on ASCL1 briefly, open closed DNA, and guide these cells toward photoreceptors so vision may improve.
Take‑home: Transient mRNA + epigenetic priming + fate factors + ocular LNP delivery.
Executive Summary
- Problem: DR neurodegeneration; MG in mammals rarely regenerate.
- Approach: ASCL1‑mRNA + HDACi; add CRX/OTX2/NRL/NEUROD1 for photoreceptor fate.
- Delivery: Intravitreal LNPs tuned for MG uptake/endosomal escape.
- Outcomes: OCT/ERG improvement with safety suitable for FIHT.
Background
MG span the retina; in DR they turn reactive (STAT3↑, GFAP↑). Early neuronal damage precedes vascular signs.
Evidence Map
- Zebrafish: robust MG regeneration.
- Mouse: ASCL1 + epigenetic support induces neurogenesis (debated durability).
- Human MG/organoids: progenitor features; rod‑like generation ex vivo.
Mechanisms
- ASCL1 pioneer factor; chromatin opening via HDACi.
- CRX/OTX2 commit fate; NRL/NR2E3 push rods; NEUROD1 maturation.
- Notch/STAT3 modulate gliosis.
mRNA/LNP Platform
| Component | Notes |
|---|---|
| mRNA | Cap‑1, modNTs, optimized UTRs; transient days. |
| LNP | Ionizable lipid + helper lipids; tune pKa, PEG. |
| Targeting | Ligands/promoter strategies; intravitreal. |
| Escape | pH‑responsive lipids aid endosomal escape. |
Preclinical Plan
- In vitro: MG/organoids → ATAC‑seq, RNA‑seq.
- Ex vivo: human explants.
- In vivo: DR rodents → OCT/ERG/markers.
- Safety: proliferation, gliosis, immunogenicity.
Clinical Development
- Ph1/2a: dose‑ascending intravitreal; AEs, BCVA, ERG, OCT.
- Ph2b: randomized vs best care; functional signal + safety.
Manufacturing (CMC)
- mRNA: dsRNA removal, identity/purity, potency.
- LNP: size/DLS, PDI, ζ, encapsulation %, sterility.
- Fill/finish: aseptic unit‑dose, CCI, E/L (ocular).
Risks & Mitigations
- Low conversion → multi‑factor pulses.
- Gliosis → modulate STAT3/Notch/inflammation.
- Ocular safety → titrate dose, prophylaxis.
FAQs
Will this replace anti‑VEGF?
No; may complement it.
Why mRNA?
Transient, tunable, non‑integrating.
What proves success?
OCT/ERG + markers + connectivity + safety.
Poster
Place your PDF next to this file as poster.pdf.
Quiz (Sample)
Score: 0 / 0
ASCL1 is…
A pioneer transcription factor
An ion channel
A lipid
References
- 1. Jorstad NL, et al. 2017 Nature.
- 2. Todd L, et al. 2021 Cell Reports.
- 3. Pollak J, et al. 2013 Development.
- … up to 35.
Authors & Contact
Authors: Meet Yashraj Singh, Sakshi, Paras Goyal
Mentor: Mrs. Pratibha Sharma
Affiliation: UIPS, Panjab University, Chandigarh
Emails: meet@smayan.in • sakshi21524@gmail.com • parasgoyal1616@gmail.com