AA AAV9. 9. L LAM AMP-2B R 2B Reverses M Metabol olic a and - - PowerPoint PPT Presentation

AA AAV9.

• 9. L

LAM AMP-2B R 2B Reverses M Metabol

• lic a

and Ph Physiologic Mu Multiorgan Dy Dysfunc uncti tion n in n a Mur urine ne Mo Model el of Da Dano non Di Disease

Ana Maria Manso, Emily Gault, Sherin Hashem, Bradley Nelson, Elizza Villarruel, Pavan Battiprolu, Annahita Keravala, Yusu Gu, Nancy Dalton, Kirk Hammond, Kirk Peterson, Eric Adler

UC UCSD School

• ol of
• f Medicine

Di Disclosur ure Informa mati tion

• Ana Maria Manso is a consultant for Rocket Pharmaceuticals.
• Pavan Battiprolu and Annahita Keravala are employees of Rocket Pharmaceuticals
• Eric Adler is a shareholder of Rocket Pharmaceuticals.

Da Dano non Di Disease: : Clini nical Mani nifestati tions ns

• Lethal

X-linked disorder caused by mutations in the Lys ysosomal Asso ssociated Mem embrane Pro rotein 2 (LA LAMP-2) gene, a lysosomal pr protein cr criti tical fo for au autophag agy.

• Ca

Cardi diomyopa path thy, sk skeletal my myopathy, an and in intelle llectual al di disabi bility ty.

• Other less prevalent symptoms include retinal disease,

hepatic disease, and pulmonary disease.

• Patients develop se

severe hy hypertrophic ca cardiomyopathy that progresses to heart failure

• Mo

Most pa pati tients ts di die in in the their 20 20-30 30s wit without he heart tr trans nspl plantati tion

• Affected males have the most severe phenotype, some

heterozygous females will also show evidence of disease.

LC3 II LC3 II LC3II LC3 II LC3 II LC3 II LAMP-2 LC3 II LC3 II LC3 II LAMP-2 LAMP-2

Au Autop

• phagy

gy

Impa Impaired ed au autophag agosome-ly lysosome fu fusio ion im impair airs au autophag agy flu flux in in Da Danon di disea ease e

Da Danon Di Disease

LA LAMP-2 i 2 isof

• for
• rms

Chi et al., 2018

LA LAMP-2B 2B is s the predominant iso soform expresse ssed in ca cardiomyocytesan and has as been postula lated to be cr critical in the pathogenesis of Da Danondi disea ease. e.

Aim: Evaluate the efficacy y of gene therapy y with adeno- as associa iated d vir viral al 9 (AA 9 (AAV9-LA LAMP-2B 2B) v ) vector

• r i

in a a m mou

• use

mo mode del of Da Dano non dis diseas ase.

AA AAV9-LA LAMP-2B 2B treatment in in LA LAMP-2 KO KO mice study

√ √

12 weeks 24 weeks

Ad Administration

• n of
• f AA

AAV9. 9.LAM AMP2B 2B showed ed dos

• se-de

depe pende ndent expr pressio ion n of mR mRNA NA and and human human LAMP-2B 2B pr protein in in in he hear art tis issue ue from m LA LAMP2 KO KO mice together with an improvement in aut autophag phagic ic fl flux (L (LC3 C3 II II level els) 24 24 weeks pos

• st-inj

injectio ion

LC3 II LC3 II LC3II LC3 II LC3 II LC3 II LC3 II LAMP-2

#P<0.05, ##P<0.01, ###P<0.001 vs PBS *P<0.05, **P<0.01, ***P<0.001, ****P<0.001 vs WT ^P<0.05, ^^^^P<0.001 vs 1e13

AA AAV9. 9.LAM AMP-2B 2B administration

• n was assoc
• ciated with an improvement in the accumulation
• n of
• f

aut autophag phagic ic st structures in hearts of LA LAMP-2 2 KO KO mice.

Ca Cardiac con

• ntractility and relaxation
• n were also
• improved in a dos
• se-de

depe pende ndent manne manner in in the he AA AAV9. 9.LAM AMP2B 2B trea eated ed LA LAMP2 KO KO mice compared to PBS controls 24 24 weeks pos

• st-inj

injectio ion

#P<0.05, ##P<0.01, ###P<0.001 vs PBS **P<0.01, ***P<0.001, ****P<0.001 vs WT

Ca Cardiac Co Contractility Ca Cardiac Relaxation

Ad Administration

• n of
• f AA

AAV9. 9.LAM AMP-2B 2B showed dos

• se-de

depe pende ndent expr pressio ion n of mR mRNA NA and and human human LA LAMP-2B 2B prot

• tein in liver tissue fr

from

• m LA

LAMP-2 2 KO KO mice together with an improvement in aut autophag phagic ic fl flux (L (LC3 C3 II levels)

#P<0.05 vs PBS **P<0.01, vs WT

AA AAV9. 9.LAM AMP2B 2B administration

• n was assoc
• ciated

ed with an improvem emen ent in the e accumulation

• n of
• f

aut autophag phagic ic st structures in livers of LA LAMP-2 2 KO KO mice.

AA AAV9. 9.LAM AMP-2B 2B administration

• n was assoc
• ciated with an improvement in the serum levels of
• f AL

ALP and AL ALT in LA LAMP-2 2 KO KO mice.

Al Alkaline phos

• sphatase

Al Alanine am amin inotran ansferas ase

#P<0.05, ##P<0.01, ####P<0.0001 vs PBS **P<0.01, ****P<0.001 vs WT

Ad Administration

• n of
• f AA

AAV9. 9.LAM AMP-2B 2B showed dos

• se-de

depe pende ndent expr pressio ion n of human human LAMP-2B 2B prot

• tein in

sk skeletal musc scle tissu ssue fr from

• m LA

LAMP-2 2 KO KO mice together with an improvement in aut autophag phagic ic fl flux (L (LC3 C3 II levels)

#P<0.05, ##P<0.01 vs PBS *P<0.05 ,**P<0.01 vs WT

AA AAV9. 9.LAM AMP-2B 2B administration

• n was assoc
• ciated with an improvement in the accumulation
• n of
• f

aut autophag phagic ic st structures in sk skeletal musc scle of LA LAMP-2 2 KO KO mice.

Su Summary

• Administration of AAV9.LAMP-2B showed dose-dependent expression of human LAMP-2B

transcript and protein in heart, liver and skeletal muscle.

• AAV9.LAMP-2B administration was associated with an improvement in autophagic flux (LC3 II

levels) as well as in the accumulation of autophagic structures in all three tissue.

• Cardiac function as well as hepatic damage were also improved in the AAV9.LAMP-2B treated

LAMP-2 KO mice compared to PBS controls

• Concl

clusion: Th These da data in indic icate th that AA AAV9.LAM LAMP-2B ge gene tr transfer im improves th the me metabolic an and ph physiologic mu multi tiorgan dysfunct ction in in th the mo mouse mo model of

• f Da

Danon di disease an and de demons nstrate pe persistent tr treatme tment effect cts up up to to 6-mo month ths po post inject ction, in indic icatin ing th the dur durabi bility of

• f th

the th therapy.

Acknowledgments

• Dr. Eric Adler

Emily Gault Elizza Villarruel Yusu Gu Paul Bushway Angel Soto-Hermida Chao Chen Sherin Hashem Bradly Nelson Michela Brambatti Pavan Battiprolu Annahita Keravala Jonathan Schwartz Gaurav Shah

• H. S. Lopez Family Foundation founded

by Czarina and Humberto S. Lopez