Systemic administration of exosomal siRNA inhibited huntingtinexpression in transgenic mice of Huntington's disease
PEI, Zhong, MD & PhD Department of NeurologyThe First Affiliated Hospital Sun Yat-Sen UniversityGuangzhou, China
Peter Como HD Research Symposium at HSG 2019: Navigating HD
Sacramento, CANov 7, 2019
Outline
p HD in China
Chinese Huntington’s Disease Network (CHDN)
p Exosomal siRNA-mediated inhibition of HTT
2009-20103 exploratory workshops
2011Official Launch
2013 Established 11 Centers in China
2014Published First Collaborative Article
ShenyangBeijing(2)
SuzhouShanghaiHangzhouFuzhou
Guangzhou
ChangshaChengdu Wuhan
Urumqi Lanzhou
Nanning
ShenyangBeijing(2)
SuzhouShanghaiHangzhouFuzhou
Guangzhou
ChangshaChengdu Wuhan
Urumqi Lanzhou
Nanning
Committed CentersPotential Centers
2011 Launch Meeting
Prof. Jean-marc Burgunder
Huntingtin gene CAG repeat numbers in Chinese patients with Huntington’s disease and controls
Chinese Huntington’s Disease network (CHDN)
Year年份
Pedigree家系
Patient Total确诊患者
2013 457 660
2014 497 733
2015 575 896
2016 634 955
2019 891 1454
The number of patients registered in CHDNCHDN中心收录患者总人数统计
Category Patient Total确诊患者
Persons at risk (blood relative)潜在患者(有血缘关系的亲属)
No. Approx.4000~7000
Approx.15000~30000
Estimated HD Patients in China中国HD患者预估人数
*Estimation based on prevalence rate in published articles from Hongkong and Taiwan (2~4/106), actual number may be higher
Steering Committee• Call for PI Meeting• Decision making• Drawing up project proposal
Central Coordination• Information gathering and distribution
• Communication central• Meeting and training organization
• Project coordination
Local CentersDaily patient care
• Conduct individual and collaborative research
Patient Advocacy• Bridge between
HD patients and researchers
• Promoting awareness of HD
Patients & Families
Health Professionals
Honorary Members• Advisory to SC• Esteemed Researches
in the field
CHDN: Administration and Management
Multi-center clinical observation study
• General Clinical Forms
Stroop Test, Symbol Digit Modality Test, Trail-making Test MMSE, Verbal Fluency Test
UHDRS: Total Function Capacity, Functional Assessment ScaleIndependence Scale
UHDRS Behavior, PBA-s, BDI II, HAD, SIS
History (medical, disease, psychiatric, family)Scocio-demographic (Fixed & Variable)Pseudonymisation, Informed ConsentCurrent TherapiesHDCC (On-set assessment)General neurological assessments (for normal control)
• Functional
• Behavioral
• Cognitive
CHDN Clinical Activity
CHDN Physician Training
No. of people累积培训人数
Year年份
Local Training中心培训
Training Session
CHDN培训
PBA training by EHDN
EHDN培训
Chinese-PBA training
PBAs中文培训
EHDN assoc.member
EHDN附属会员2010-2012
15~20 each time - - - 3
2013
-
30 - - 11
2014 40 2 6 44
2015 50 2 18 51
2016 57 2 25 53
2017 65 2 33 53
CHDN Clinical Activity
Training & Certification
On-site Training
Certification
2012 Local Training in Guangzhou Center
EHDNCHDN
Exchange fellowship
CertificationTwo way academic exchange
Ensure only qualified physicians and researchers can participate
CHDN Activity
Training & CertificationCHDN Clinical Activity
Annual PI meetings
Guangzhou广州
Hangzhou杭州
Chengdu成都
Beijing北京
Fuzhou福州
CHDN Clinical Activity
Chinese HD Association
President :Xi Cao
Contact us:
Provide Support Network
for Patients and Families
Advocate for Better Policies for HD Community
Vice President
international huntington association
Since 2013
CHDN Activity
Survey on HD Patients' Living Conditions in China
1. Lack of HD knowledge and disease management
37.25% of the patients never received any treatment for HD.Only 1.96% of the patients had surgical treatment for HD and 0.98% had physiotherapyDBS procedure is not covered by local medical insurance inmost cities
2. Inadequate medical insurance system
17.96% of the patients in the survey don't have any medical insuranceHD is not covered by any commercial insurance plans Many HD families have more than one HD patient, resulting in decreasing family income, a lot of families have to live on social benefits, which barely cover their expenses.
Chinese HD Association CHDN Activity
CHDN Translational Research Collaboration
IPS Cells
Animals
Isolating epithelial cells
Reprogramming the cells into UiPSCs
Patient-specific cell model
Getting urine from patients
PotentialTreatment
Drug Testing
Understand pathology
Guangzhou center in collaboration with South China Institute for Stem Cell Biology and Regenerative Medicine
HD Transgenic Pig HD Knock-In Pig(Unpublished)
PIG
MICE Novel Full-length Htt Mouse ModelBy Prof. Li Chenjian’s team at Beijing University
By Prof. Li Xiaojiang & Prof. Li Shihua’s team& Prof. Lai Liangxue’s team at Chinese Academy of Science
Potential Imaging
Studies
Future Collaborations
test IPS treatment etc.
CHDN Activity
Preclinical model: Huntingtin knockin pig (Cell 173, 1-14, May 3, 2018)Li Xiao-Jiang Lai Liangxue
CHDN Translational Research Collaboration
Exosomal siRNA-mediated inhibition of HTT in vivo
Chen Xi Department of Life sciences Nanjing University
PEI, ZhongDepartment of Neurology The First Affiliated Hospital Sun Yet-Sen University
CHDN Translational Research Collaboration
Wild EJ et al, Lancet Neurol, 2017
siRNA and Huntingtin-lowering therapeutics
Systemic siRNA delivery and tissue targeting
Hartmann G, et al Nature Biotechnology 2011
Barriers:q Unfavorable chemical properties
Negative charges etc
q Instability
q Plasma half-lives (10 min)
Big challenges for siRNA therapeutics
p Exosomes:
ü Extracellular membrane vesicles (30-150 nm).ü Transmission of macromolecules between cells.
Advantages as drug vehicles:ü Small sizeü Low immunogenicityü Long half-life in the circulation ü Ability to load various cargoes ü Easiness to cross Brain-blood barrier
Exosomes: a novel biocarrier for siRNA Delivery to the brain
Zhang M. et al, J. Mater. Chem. 2019
Efficient scalable production of clinical-grade exosomes
Brain specific delivery
Bottleneck 1 Bottleneck 2
Challenges for exosome-based therapeutics in CNS diseases
Liver: the endogenous exosome factory for siRNA
q siRNA accumulated in liver following I.V. injection of plasmids
q The liver is a primary site of exosome uptake
Chen XNanjing University
Liu et al in preparation
Large production of exosomes
Self-assembly exosomal siRNA delivery system
RVG-Lamp2b modified exosomes: Brain specific delivery
q RVG (rabies virus glycoprotein) CNS-specific uptake of exosomes
q Lamb2b (lysosomal associated membrane protein-2b)Link RVG to the external exosomal surface
RVG
RVGLamb2b
Brain cells
RVG/LAMP2b significantly increased the delivery of siRNA to the brain
b rain
live r
s p lee n
lun g
k idn e y
02 0 04 0 06 0 08 0 0
1 0 0 03 0 0 0
3 5 0 0
4 0 0 0
siR
NA
le
ve
l(c
op
y/c
ell
)
P B SS c ra m b les iR N AR V G -s iR N A
UD
UD
UD
UD
UD
UD
UD
UD
UD
UD
UD
siRNAScramble RVG-siRNA
Liver
Brain
Lung
In situ hybridization
Chen XNanjing University
DAPI
eGFP
RVG-siRNA
Liver Brain
siRNAScramble RVG-siRNAScramble siRNA
Cre eGFP reporter mouse
Exosomal siRNA2 and 5 exhibited potent inhibition of huntingtin expression in SHSY5Y cells and BACHD mice
SHSY5Y cells
Intravenous administration of exosomal siRNA for two weeks reduced huntingtin expression in N171-82Q mice
YAC 1286weeks
Plasmid injection10mg/kg iv. Twice a week.
RotarodOpen Field
1week
Exosomal siRNA for 8 weeks significantly suppressed huntingtin expression in both cortex and striatum in YAC128 mice
YAC 1286weeks
Plasmid injection10mg/kg iv. Twice a week.
RotarodOpen Field
1week
Intravenous administration of exosomal siRNA for 8 weeks significantly improved motor function in YAC128 mice
Exosomal siRNA reduced HTT aggregates in YAC128 mice
Normal levels of Liver enzymes
Preliminary data: Systemic exosomal siRNA did not induced toxicity in mice
Normal morphology of major organs
Summary
• Systemic exosomal siRNA lowers HTT and improves disease-phenotypes.
• Great potential for an affordable, non-invasive solution for HTT lowering therapy.
AcknowledgmentsHD clinic
Chen DBYou HQWu QLiang XL
Exosomal siRNAChen X’s lab Liu JYu MJNanjing University
Pei’ labWu TT Chinese Huntington’s Disease Network
Jean-Marc BurgunderShang HuiFangCHDI foundationEHDN
Christopher A. RossWanli W. SmithJohns Hopkins University
Thanks
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