Cross-species RNAi therapy via AAV delivery alleviates neuropathic pain by targeting GCH1

Tetrahydrobiopterin (BH4) expression is normally strictly controlled; however, its intracellular levels increase considerably following nerve damage. GTP cyclohydrolase I (GCH1) plays a crucial role in regulating BH4 concentration, with an upregulation observed in the dorsal root ganglion in cases o...

Full description

Saved in:
Bibliographic Details
Main Authors: Heesue Chang, Kyoung Jin Lee, Minkyung Park, Ha-Na Woo, Ji Hyun Kim, Im Kyeung Kang, Hyochan Park, Chan Hee Chon, Heuiran Lee, Hyun Ho Jung
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:Neurotherapeutics
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S1878747924001983
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850180998867714048
author Heesue Chang
Kyoung Jin Lee
Minkyung Park
Ha-Na Woo
Ji Hyun Kim
Im Kyeung Kang
Hyochan Park
Chan Hee Chon
Heuiran Lee
Hyun Ho Jung
author_facet Heesue Chang
Kyoung Jin Lee
Minkyung Park
Ha-Na Woo
Ji Hyun Kim
Im Kyeung Kang
Hyochan Park
Chan Hee Chon
Heuiran Lee
Hyun Ho Jung
author_sort Heesue Chang
collection DOAJ
description Tetrahydrobiopterin (BH4) expression is normally strictly controlled; however, its intracellular levels increase considerably following nerve damage. GTP cyclohydrolase I (GCH1) plays a crucial role in regulating BH4 concentration, with an upregulation observed in the dorsal root ganglion in cases of neuropathic pain. In this study, we aimed to develop and evaluate the clinical potential of an RNA interference-based adeno-associated virus (AAV) targeting GCH1 across various species to decrease BH4 levels and, consequently, alleviate neuropathic pain symptoms. We identified universal small-interfering RNA sequences effective across species and developed an AAV-u-shRNA that successfully suppressed GCH1 expression with minimal off-target effects. Male Sprague Dawley rats were divided into four groups: normal, spared nerve injury, AAV-shCON, and AAV-u-shGCH1. The rats were sacrificed on post-injection day 28 to collect blood for BH4 level assessment. The AAV-u-shGCH1 group demonstrated remarkable improvement in the mechanical withdrawal threshold by PID 28, significantly outperforming the normal, spared nerve injury, and AAV-shCON groups. Plasma BH4 levels confirmed that AAV-u-shGCH1 effectively reduced neuropathic pain by inhibiting BH4 synthesis in vivo, introducing a novel, multispecies-compatible therapeutic strategy. Our results suggest that a single application of AAV-u-shGCH1 could offer a viable solution for neuropathic pain relief.
format Article
id doaj-art-8a8a78fc47534766b452819ae3c1caff
institution OA Journals
issn 1878-7479
language English
publishDate 2025-03-01
publisher Elsevier
record_format Article
series Neurotherapeutics
spelling doaj-art-8a8a78fc47534766b452819ae3c1caff2025-08-20T02:18:00ZengElsevierNeurotherapeutics1878-74792025-03-01222e0051110.1016/j.neurot.2024.e00511Cross-species RNAi therapy via AAV delivery alleviates neuropathic pain by targeting GCH1Heesue Chang0Kyoung Jin Lee1Minkyung Park2Ha-Na Woo3Ji Hyun Kim4Im Kyeung Kang5Hyochan Park6Chan Hee Chon7Heuiran Lee8Hyun Ho Jung9Department of Neurosurgery, Yonsei University College of Medicine, Seoul, Republic of KoreaDepartment of Microbiology, University of Ulsan College of Medicine, Seoul, Republic of Korea; Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Seoul, Republic of KoreaDivision of Systems Neuroscience, Department of Psychiatry, Columbia University Irving Medical Center (CUIMC), and Research Foundation for Mental Hygiene, Inc. (RFMH), New York State Psychiatric Institute (NYSPI), New York, NY, 10032, USABio-Medical Institute of Technology, University of Ulsan College of Medicine, Seoul, Republic of Korea; Department of Biochemistry & Molecular Biology, University of Ulsan College of Medicine, Seoul, Republic of KoreaDepartment of Microbiology, University of Ulsan College of Medicine, Seoul, Republic of Korea; Bio-Medical Institute of Technology, University of Ulsan College of Medicine, Seoul, Republic of KoreaDepartment of Microbiology, University of Ulsan College of Medicine, Seoul, Republic of Korea; Asan Medical Institute of Convergence Science and Technology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of KoreaInventage Lab, Seongnam, Republic of KoreaInventage Lab, Seongnam, Republic of KoreaBio-Medical Institute of Technology, University of Ulsan College of Medicine, Seoul, Republic of Korea; Department of Microbiology, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Republic of Korea; Corresponding authors.Department of Neurosurgery, Yonsei University College of Medicine, Seoul, Republic of Korea; Department of Neurosurgery, Brain Research institute, Yonsei University College of Medicine, Seoul, Republic of Korea; Corresponding authors.Tetrahydrobiopterin (BH4) expression is normally strictly controlled; however, its intracellular levels increase considerably following nerve damage. GTP cyclohydrolase I (GCH1) plays a crucial role in regulating BH4 concentration, with an upregulation observed in the dorsal root ganglion in cases of neuropathic pain. In this study, we aimed to develop and evaluate the clinical potential of an RNA interference-based adeno-associated virus (AAV) targeting GCH1 across various species to decrease BH4 levels and, consequently, alleviate neuropathic pain symptoms. We identified universal small-interfering RNA sequences effective across species and developed an AAV-u-shRNA that successfully suppressed GCH1 expression with minimal off-target effects. Male Sprague Dawley rats were divided into four groups: normal, spared nerve injury, AAV-shCON, and AAV-u-shGCH1. The rats were sacrificed on post-injection day 28 to collect blood for BH4 level assessment. The AAV-u-shGCH1 group demonstrated remarkable improvement in the mechanical withdrawal threshold by PID 28, significantly outperforming the normal, spared nerve injury, and AAV-shCON groups. Plasma BH4 levels confirmed that AAV-u-shGCH1 effectively reduced neuropathic pain by inhibiting BH4 synthesis in vivo, introducing a novel, multispecies-compatible therapeutic strategy. Our results suggest that a single application of AAV-u-shGCH1 could offer a viable solution for neuropathic pain relief.http://www.sciencedirect.com/science/article/pii/S1878747924001983Neuropathic painAdeno-associated virusTetrahydrobiopterinGTP cyclohydrolase IRNA interferenceMultispecies-compatibility
spellingShingle Heesue Chang
Kyoung Jin Lee
Minkyung Park
Ha-Na Woo
Ji Hyun Kim
Im Kyeung Kang
Hyochan Park
Chan Hee Chon
Heuiran Lee
Hyun Ho Jung
Cross-species RNAi therapy via AAV delivery alleviates neuropathic pain by targeting GCH1
Neurotherapeutics
Neuropathic pain
Adeno-associated virus
Tetrahydrobiopterin
GTP cyclohydrolase I
RNA interference
Multispecies-compatibility
title Cross-species RNAi therapy via AAV delivery alleviates neuropathic pain by targeting GCH1
title_full Cross-species RNAi therapy via AAV delivery alleviates neuropathic pain by targeting GCH1
title_fullStr Cross-species RNAi therapy via AAV delivery alleviates neuropathic pain by targeting GCH1
title_full_unstemmed Cross-species RNAi therapy via AAV delivery alleviates neuropathic pain by targeting GCH1
title_short Cross-species RNAi therapy via AAV delivery alleviates neuropathic pain by targeting GCH1
title_sort cross species rnai therapy via aav delivery alleviates neuropathic pain by targeting gch1
topic Neuropathic pain
Adeno-associated virus
Tetrahydrobiopterin
GTP cyclohydrolase I
RNA interference
Multispecies-compatibility
url http://www.sciencedirect.com/science/article/pii/S1878747924001983
work_keys_str_mv AT heesuechang crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT kyoungjinlee crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT minkyungpark crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT hanawoo crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT jihyunkim crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT imkyeungkang crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT hyochanpark crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT chanheechon crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT heuiranlee crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1
AT hyunhojung crossspeciesrnaitherapyviaaavdeliveryalleviatesneuropathicpainbytargetinggch1