In vivo self-assembled siRNAs within small extracellular vesicles attenuate LRRK2-induced neurodegeneration in Parkinson's disease models

Li Zhang; Penglu Chen; Tiantian Chen; Lishan Lin; Xiaoyi Ji; Jin Liu; Heng Huang; Tanvikhaa Saravanan; Hannah Fuehrer; Christopher A Ross; Wanli W Smith; Zhong Pei; Xi Chen

doi:10.1016/j.jconrel.2024.12.045

In vivo self-assembled siRNAs within small extracellular vesicles attenuate LRRK2-induced neurodegeneration in Parkinson's disease models

J Control Release. 2024 Dec 23:S0168-3659(24)00900-3. doi: 10.1016/j.jconrel.2024.12.045. Online ahead of print.

Authors

Li Zhang¹, Penglu Chen², Tiantian Chen², Lishan Lin³, Xiaoyi Ji⁴, Jin Liu², Heng Huang⁵, Tanvikhaa Saravanan⁶, Hannah Fuehrer⁶, Christopher A Ross⁷, Wanli W Smith⁸, Zhong Pei⁹, Xi Chen¹⁰

Affiliations

¹ Nanjing Drum Tower Hospital Center of Molecular Diagnostic and Therapy, State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology, NJU Advanced Institute of Life Sciences (NAILS), School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210023, China; Department of Pharmacology, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, China.
² Nanjing Drum Tower Hospital Center of Molecular Diagnostic and Therapy, State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology, NJU Advanced Institute of Life Sciences (NAILS), School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210023, China.
³ Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Diagnosis and Treatment of Major Neurological Diseases, National Key Clinical Department and Key Discipline of Neurology, Guangzhou, Guangdong 510080, China; Departments of Psychiatry and Pharmacology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
⁴ Department of Pharmacology, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210023, China.
⁵ Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Diagnosis and Treatment of Major Neurological Diseases, National Key Clinical Department and Key Discipline of Neurology, Guangzhou, Guangdong 510080, China.
⁶ Departments of Psychiatry and Pharmacology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
⁷ Departments of Psychiatry and Pharmacology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA; Departments of Neuroscience and Neurology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
⁸ Departments of Psychiatry and Pharmacology, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA. Electronic address: wsmith60@jhmi.edu.
⁹ Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Diagnosis and Treatment of Major Neurological Diseases, National Key Clinical Department and Key Discipline of Neurology, Guangzhou, Guangdong 510080, China. Electronic address: peizhong@mail.sysu.edu.cn.
¹⁰ Nanjing Drum Tower Hospital Center of Molecular Diagnostic and Therapy, State Key Laboratory of Pharmaceutical Biotechnology, Jiangsu Engineering Research Center for MicroRNA Biology and Biotechnology, NJU Advanced Institute of Life Sciences (NAILS), School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210023, China; The Second People's Hospital of Changzhou, The Third Affiliated Hospital of Nanjing Medical University, Changzhou Medical Center, Nanjing Medical University, Changzhou, Jiangsu 213003, China. Electronic address: xichen@nju.edu.cn.

PMID: 39722304
DOI: 10.1016/j.jconrel.2024.12.045

Abstract

Rationale: Mutations in the leucine-rich repeat kinase 2 (LRRK2) gene play an important role in Parkinson's disease (PD) pathogenesis, and downregulation of LRRK2 has become a promising therapy for PD. Here, we developed a synthetic biology strategy for the self-assembly and delivery of small interfering RNAs (siRNAs) of LRRK2 into the substantia nigra via small extracellular vesicles (sEVs) using a genetic circuit (in the form of naked DNA plasmid) to attenuate PD-like phenotypes in mouse model.

Methods: We generated the genetic circuit encoding both a neuron-targeting rabies virus glycoprotein (RVG) tag and a LRRK2 siRNA under the control of a cytomegalovirus (CMV) promoter, and assessed its therapeutic effects using LRRK2^R1441G mouse models of PD.

Results: After intravenous injection, the genetic circuit was taken up by the host liver to reprogram liver cells to produce and self-assemble LRRK2 siRNAs into sEVs, and then the sEV-enclosed LRRK2 siRNAs were further transferred by the endogenous circulating system of sEVs and guided by the RVG tag to the substantia nigra. Intravenous injection of this genetic circuit reduced total and phosphorylated LRRK2 levels in the substantia nigra, and attenuated PD-like phenotypes in two mouse models by rescuing LRRK2^R1441G-induced dopaminergic neurodegeneration and reducing microgliosis.

Conclusion: This study provides an efficient therapeutic strategy to attenuate LRRK2-induced neurodegeneration and neuroinflammation in PD mouse models, and may open a new avenue to safely deliver siRNA into brain after peripheral intravenous injection and facilitate PD treatment.

Keywords: Extracellular vesicle; In vivo self-assembled siRNA; LRRK2; Neurodegeneration; Parkinson's disease.