# Gene Therapy Targeting of CNTFRalpha and CLC in Muscle to Treat ALS

> **NIH NIH R01** · UNIVERSITY OF CINCINNATI · 2022 · $405,917

## Abstract

Project Summary: Muscle ciliary neurotrophic factor receptor α (CNTFRα) expression is induced: 1) by
denervating nerve lesion53-55, 2) in human denervating diseases56,57, including ALS56, and 3) in all ALS models
tested. Muscle CNTFRα knockdown inhibits motor neuron (MN) axon regeneration and motor recovery after
nerve lesion53 and accelerates disease in all the ALS models, suggesting the muscle CNTFRα induction is a
neuroprotective response that could be enhanced to treat ALS. We increased muscle CNTFRα in SOD1G93A
ALS mice with an AAV vector (AAV1.1-CNTFRα), extending survival and increasing motor function (without
side effect) even when started well after symptom onset, making it arguably the most clinically promising
treatment to date since human ALS is not treated until well after symptom onset58-61. We find vector-derived
CNTFRα protein translocated to MNs and increased MN terminals, suggesting a mechanism of action. This
treatment should: 1) inhibit most/all ALS, 2) work independent of ALS causes, and 3) be translatable since
muscle expression can be increased with approved human gene therapy techniques42-47,49,50, like those we use
here. The ligand most likely involved in muscle CNTFRα's anti-ALS effects is muscle cardiotrophin-like
cytokine (CLC). We similarly increased muscle CLC in SOD1G93A mice, again extending survival without side
effect, making it another very promising new ALS treatment. Combining the two treatments suggests this could
be an even more effective treatment. We will: Aim 1: Optimize and Characterize muscle CNTFRα
enhancement as an ALS treatment. To maximize effect, we will dose response test in SOD1G93A mice: 1) a
codon optimized CNTFRα cDNA, 2) an AAV capsid with greatly enhanced muscle transduction (AAV1.1), 3) a
muscle specific promotor (tMCK), 4) self-complementary AAV for faster and greater expression, and 5) IV
injection of another next-gen capsid (AAV2i8) for transduction of more muscles. The best treatment will be
further examined with: 1) rotarod and grip strength tests, 2) qRT-PCR and in situ hybridization for vector-
derived CNTFRα RNA, 3) HA tag anatomy to localize vector-derived CNTFRα protein and identify potential
sites of action, 4) multi-label immunohistochemistry to determine effects on ALS degeneration, and 5) two
other diverse ALS models (SOD1G37R and TDP-43Q331K mice) to broadly test the treatment. Aim 2: Optimize
and Characterize muscle CLC enhancement as an ALS treatment. We will run experiments exactly as in
Aim 1, except with CLC instead of CNTFRα. Aim 3: Optimize and Characterize combined CNTFRα and
CLC treatment. Muscle CLC and CNTFRα are likely released as a MN protective CLC/CNTFRα complex
such that an increase in both CLC and CNTFRα may further enhance efficacy. A pilot with a 1:1 ratio of the
non-optimized CLC and CNTFRα vectors found a substantially enhanced effect in females. We will test other
ratios with CNTFRα and CLC treatments optimized in Aims 1 and 2, to further increase the fe...

## Key facts

- **NIH application ID:** 10427242
- **Project number:** 5R01NS113917-04
- **Recipient organization:** UNIVERSITY OF CINCINNATI
- **Principal Investigator:** Alexander John MacLennan
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $405,917
- **Award type:** 5
- **Project period:** 2019-09-15 → 2024-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10427242

## Citation

> US National Institutes of Health, RePORTER application 10427242, Gene Therapy Targeting of CNTFRalpha and CLC in Muscle to Treat ALS (5R01NS113917-04). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10427242. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*