Characterization of Niemann-Pick Type C mutant mice using automated cage monitoring systems

Transcription

1 Characterization of Niemann-Pick Type C mutant mice using automated cage monitoring systems Dr. Sangeetha V. Iyer Perlara PBC San Francisco, CA sangeetha@perlara.com

2 Contents Introduction to Perlara PBC Background of Niemann Pick Type C disease Prevailing literature Introduction to Vium Inc. Results Summary Future Direction

3 Introduction to Perlara PBC PERLARA IS A SCIENTIFIC DISCOVERY PUBLIC BENEFIT CORPORATION Perlara is a global patient portal and drug discovery platform for families affected by rare disease - 1 in 10 on Earth.

4 Perlara s Model Patient families and advocacy groups initiate PerlQuests through Perlara Perlara uses its novel platform of engineered model organisms to screen for therapeutics against patient disease Perlara and patient groups partner with BioPharma and to advance precision drug candidates to the clinic.

5 Perlara s Platform Approach Parallel screens in animals yield orally bioavailable leads Patient-derived cells for mutation-specific in vitro validation Fast path to mouse tox and efficacy studies Powerful genetic toolkits for target ID (RNAi)

6 Rationale Rare diseases associated with a conserved, single gene Identify prevalence of mutational variants Model mutations in simple organisms that have conserved genes Proof of Concept studies C G G T A C AC G G T A C T C G G T A C C G G T A C A A spectrum of mutations associated with each rare disease Cellular phenotypes can be different Reverse phenotype in model organisms and patient-derived cells Precision drugs to market

7 Niemann Pick Type C Disease is a hereditary lysosomal storage disorder LDL Wildtype LDL npc1 NPCI gene codes for a protein, required for cholesterol trafficking in absence of npc1 cholesterol accumulates in lysosomes NPC mortality: teenage years to early adulthood NPC Disease: ~200 US patients No FDA approved therapeutic NPC1 gene homologs are present in Yeast, C.elegans, Drosophila and Mice.

8 We engineered NPC disease in model organisms and conducted discovery screens Phenotype Assay ($$) Throughput Slowed growth and fewer progeny Drugs that increase growth and reproduction 20K compounds/month Phenotype Assay ($$) Throughput Growth arrest as young larvae and death Suppression of growth arrest and death 10K compounds/month Phenotype Assay ($$$$) Throughput Cholesterol accumulation in endolysosomes Clearance or Redistribution of cholesterol 50K compounds/month

9 Screens results needed to be tested in preclinical model of NPC Parallel screens in animals yield orally bioavailable leads Patient-derived cells for mutation-specific in vitro validation Fast path to mouse tox and efficacy studies

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11 NPC KO mice show reduced weight gain compared to WT Voiker et al, Behavioral Brain Research, 132 (2002) 1-10

12 NPC KO mice show reduced motor abilities Open field activity Balance beam test Rotarod Voiker et al, Behavioral Brain Research, 132 (2002) 1-10

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14 Perlara s goals Establish an NPC mouse model for testing Perlara s lead candidates Determine baseline features and positive controls where appropriate Considerations include Sample size High resolution data Cost Humane treatment of animals

15 Vium Technology All standard CRO capabilities AAALAC accredited Digital Vivarium Vium data analytics Optimal Animal Environment Less human intervention Consistent measurements Fewer animals Improved Study Operation High throughput More data Data available immediately Better insights Standardized efficacy scores Novel side effect detection Can view data retrospectively Improved Decision Making More informed and earlier prioritization

16 What the Vium interface looks like

17 NPC KO animals show reduced body weight and respiration in later stages of the disease compared to WT Age (Days) Age (Days)

18 NPC KO animals show shorter lifespans compared to WT controls Age (Days) * WT s were euthanized after they outlived KO as the study was terminated

19 Time (sec.) on RR Time (sec.) on RR KO animals show a decrease in time spent on rotarod relative to WT at later stages of the disease Day 35 Day WT KO 0 WT KO

20 KO controls show a significant difference in performance on balance beam with advancing age 185 Retention Average 25 Average Sections crossed Day WT KO 0 WT KO Day WT Retention Average KO Average Sections crossed WT KO

21 Cyclo-treated KO mice live longer compared to Vehtreated controls

22 Cyclodextrin treated KO animals maintain bodyweights longer than vehicle-treated animals Weight collected manually Weight collected automatically Age (Days)

23 Cyclo-treated KO animals maintain activity levels longer, unlike vehicle-treated KO s which show decreased motion at later stages of the disease

24 Vehicle-treated KO s differ significantly on their motion in the dark cycle relative to WT and Cyclo-treated KO s

25 Vehicle-treated KO s show decreased respiration compared to WT and Cyclo-treated KO s at later stages of the disease Time (days)

26 Cyclo-treated KO s maintain their breathing rate longer compared to Veh-treated KO s independent of lightdark cycle

27 Cyclo-treated KO show differences in liver enzymes and triglycerides relative to Veh-treated KO

28 Vium s Smart Housing collects more data with less stress Conventional Method Metrics: Grip strength Locomotor activity Balance Features: Remove from cage Short duration One at a time Increased handling Increased stress Higher variability Limited data Vium Method Metrics: Motion Breathing Circadian activity Features: In-cage Continuous monitoring All animals (at scale) Objective data requires smaller n to reach significant results WT Vehicle KO Vehicle KO Cyclo Days and Treatment (P<0.0001), Days x Treatment (P<0.02) by 2-way ANOVA and Tukey Post-hoc Test

29 Summary The NPC KO line established at Vium was subjected to traditional behavioral assays such as rotarod, balance beam as well as automated recording of metrics Automated data showed trends parallel to that reported in literature and collected by traditional methods High resolution data capture using automated methods uncovered two new metrics (circadian motion and respiration) that could be exploited to benchmark disease progression and treatment Cyclodextrin, a widely-accepted positive control, showed rescue of reported disease phenotypes in NPC KO animals Other metrics not reported here include better breeding, progeny ratios, rapid response to health checks

30 Future Directions Novel mechanism Unique cell redistribution EC 50 in cells: 2µM Design dosing and experimentation paradigm for P101 Identify additional metrics to complement traditional phenotypes Blood brain barrier permeable PERL % stability in Invitro PK Assa YS 100% oral bioavailability

31 Questions?