Academic Research 2026
Academic Research / R01 Renewal

Base-Editing
Sickle Cell

A grant committee brief: reactivating fetal hemoglobin with an adenine base editor to correct sickle-cell disease without a double-strand break.

Dr. Amara Okonkwo · Children's Hospital of Philadelphia NIH NHLBI R01 · Renewal Year 1
Field Map Part One
01 · Field Map

Two therapies now treat
sickle-cell at the genome

Since 2023, Casgevy and Lyfgenia have shown ex vivo genome editing can functionally cure sickle-cell disease — but both depend on a double-strand break and myeloablative conditioning.

2026 The Gap Sickle-Cell
The Gap 03

The gap · HbF reactivation

Nuclease editing cures patients, but the double-strand break itself is now the safety question the field cannot ignore.

Okonkwo Lab · CHOP R01 HL178453
Approach 2026
Approach 04

The Approach

An adenine base editor, not a nuclease

ABE8e installs a single A>G edit at the HBG1/HBG2 −113 promoter site, recreating a naturally occurring hereditary persistence of fetal hemoglobin (HPFH) variant — no double-strand break, no cutting.

  • Delivered as mRNA + sgRNA ribonucleoprotein via electroporation into autologous CD34+ HSPCs
  • No viral vector integration — edited cells re-infused after non-genotoxic conditioning
  • Mimics a benign HPFH allele already documented in three unaffected carrier families
Image placeholder

Base-editing schematic · HBG1/HBG2 promoter locus

Okonkwo Lab · CHOP R01 HL178453
By the Numbers 2026
By the Numbers 05

What the preclinical data shows

68%

Editing efficiency achieved in CD34+ HSPCs across three independent donor lots

Sanger + deep sequencing · n=3 donors

41%

Fetal hemoglobin induced per edited allele, above the 20% threshold linked to clinical benefit

HPLC hemoglobin quantification

<0.1%

Off-target editing detected at the top 10 CIRCLE-seq nominated sites

Whole-genome + targeted amplicon sequencing

Okonkwo Lab · CHOP R01 HL178453
Study Design 2026
Study Design 06

What's inside

Four aims carry the proposal

Each aim retires a specific risk before the next begins — the design a study section expects to see.

  • Aim 1 — Optimize on-target editing and HbF induction across donor genotypes
  • Aim 2 — Xenograft safety: engraftment, off-target burden, clonal composition at 16 weeks
  • Aim 3 — GMP-compatible manufacturing and potency assay lock for IND filing
  • Aim 4 — First-in-human Phase 1 dose-and-safety trial design
  • Milestone-gated go/no-go criteria reviewed by an independent safety board at each transition
Okonkwo Lab · CHOP R01 HL178453
Standard vs Proposed 2026
Standard · Proposed 07
Current Standard Care

Allogeneic transplant or nuclease editing

Both cures that exist today carry real costs: a matched donor, or a permanent double-strand break.

  • Allo-HSCT requires a matched sibling donor — available to under 15% of patients
  • Casgevy/Lyfgenia use Cas9 nuclease cuts, with documented large-deletion and mosaic-loss events
  • Both require myeloablative busulfan conditioning
Proposed Base-Editing Approach

Autologous, cut-free, non-genotoxic

No donor search, no double-strand break, and a conditioning regimen designed to spare marrow reserve.

  • Autologous cells — every patient is their own donor
  • Base editing installs the edit without cutting both DNA strands
  • Antibody-based conditioning under evaluation to avoid busulfan toxicity
Okonkwo Lab · CHOP R01 HL178453
Investigator Voice 2026

"The mutation we're installing already exists in nature, in people who never developed sickle-cell symptoms. We're not engineering something new — we're restoring a state biology already proved was safe."

Dr. Amara Okonkwo · Principal Investigator Children's Hospital of Philadelphia, Division of Hematology
Analysis 2026
Analysis 09

Why this proposal changes the risk calculus

Significance

Sickle-cell disease affects roughly 100,000 Americans and an estimated 20 million people worldwide, disproportionately Black and of African, Middle Eastern, and South Asian descent — a population underserved by rare-disease research investment.

The two approved genome-editing therapies proved the concept but priced it near $2.2 million per patient and paired it with a genotoxic conditioning regimen many eligible patients are too sick to tolerate.

A base-editing approach that avoids double-strand breaks addresses the two objections raised most often in FDA and NIH safety reviews of first-generation gene-editing therapies: off-target genotoxicity and large structural variants.

Innovation

This is the first R01-supported program to target the HBG1/HBG2 promoter directly with a base editor rather than editing BCL11A, the erythroid enhancer targeted by Casgevy.

Because the edit recreates a naturally occurring HPFH allele, the safety argument rests on decades of clinical observation of unaffected carriers, not solely on new animal data.

The proposal is scoped narrowly: four aims, one edit, one delivery method — built to answer a reviewer's first question ("why will this work when others have tried") rather than to promise a platform.

Okonkwo Lab · CHOP R01 HL178453
Preclinical Evidence 11

Editing efficiency across three donor lots

% alleles edited · CD34+ HSPCs · preclinical data
61%
Donor A
74%
Donor B
65%
Donor C
80%
Aim 1 Target

Preclinical data · n=3 independent CD34+ HSPC donor lots · Aim 1 target shown for reference

Okonkwo Lab · 2026 11 / 16
Project Timeline 12

From bench to IND filing

01
Optimize
Screen guide RNA and editor variants across three donor genotypes to lock the lead construct (Aim 1, Months 1–12).
02
Validate
Confirm safety and engraftment in NBSGW xenograft models at 16-week endpoints (Aim 2, Months 6–24).
03
Manufacture
Transfer the locked protocol to GMP-compatible closed-system manufacturing and finalize the potency assay (Aim 3, Months 18–36).
04
File
Compile the IND package and design the Phase 1 dose-escalation trial with the study section's guidance (Aim 4, Months 30–48).
Okonkwo Lab · 2026 12 / 16
Budget 2026
Budget Allocation 13

How the $2.98M direct budget breaks down

Xenograft Safety Studies 38%
GMP Manufacturing 25%
Personnel & Core Facility Fees 22%
Sequencing & Off-Target Assays 15%

Total direct costs · 4-year R01 budget

Budget justification, Section 5.3 · figures rounded

Okonkwo Lab · 2026
Milestones 2026
Milestones 14

Four years, four go/no-go gates

Year 1
Construct lock

Guide RNA and base-editor variant selected; ≥60% editing efficiency confirmed across all three donor genotypes.

Year 2
Safety gate

16-week xenograft engraftment and off-target sequencing complete; independent safety board reviews go/no-go.

Year 3
Manufacturing lock

GMP-compatible process transferred; potency assay validated for IND-enabling studies.

Year 4
IND submission

Pre-IND meeting held with FDA; Phase 1 protocol and consent documents finalized for submission.

Okonkwo Lab · 2026 14
Risk Review 2026
Risk Review 15

How risk is reviewed, not just disclosed

01
Identify

Off-target sites nominated by CIRCLE-seq and computational prediction before any editing begins.

02
Assess

Whole-genome and targeted amplicon sequencing quantify editing at every nominated site per donor lot.

04
Monitor

An independent data and safety monitoring board reviews cumulative findings at each aim's go/no-go gate.

03
Mitigate

High-fidelity editor variants and dose titration substituted wherever off-target signal exceeds 0.1%.

Okonkwo Lab · 2026 15
Contribution 2026
Contribution 17

Contribution

How this preliminary work adds up

From bench validation to a therapy request the committee can fund

Phase 1 Trial Request
IND-Ready Manufacturing
Xenograft Safety Data
Editing Efficiency Data
Naturally Occurring HPFH Precedent
Okonkwo Lab · CHOP R01 HL178453
Next Question 2026

Next Question

What dose, in which patients, should the Phase 1 trial test first?

We are requesting a one-year no-cost extension supplement to fund a natural-history cohort of 30 pediatric and adult sickle-cell patients ahead of first-in-human dosing — a.okonkwo@chop.edu · Division of Hematology.