Corning Incorporated · NYSE:GLW
Corning — ISSCR 2023 Corning Sponsored Innovation Showcase
69 slides from Corning's conference presentation, Jun 15, 2023
Slide 1: Corning Sponsored Innovation Showcase
Corning Sponsored Innovation Showcase: Predicting Drug Responses for Conventional & IO therapies Using MicroOrganoSpheres™ Dennis Plenker, Ph.D. Xilis, Inc. iPSC-derived Vascular Cells for Vascularization of Organoids & Ischemic Tissues Ping Zhou, Ph.D. Stem Cell Program and Department of Internal M
Slide 2: Welcome and Introduction
Welcome and Introduction Catherine Siler, Ph.D. Field Application Scientist Manager (East) Corning Life Sciences
Slide 3: Today's Discussion
Today’s Discussion 3D cell culture relevance Considerations for improving 3D methods: • Standardization of methods • Increasing throughput © 2018 Corning Incorporated 3 General - Corning (L4)
Slide 4: Preclinical models for therapeutics
Preclinical models for therapeutics 2D Spheroids Organoid Mouse Ease of use Biological Complexity Throughput Human biological relevance Standardized methods Higher throughput General - Corning (L4)
Slide 5: 3D Cell Culture serves multiple disciplines
3D Cell Culture serves multiple disciplines Disease Modeling Ping Zhou, Ph.D. Dennis Plenker, Ph.D. Regenerative Precision Medicine Medicine Drug Testing iPSC-derived Vascular Cells for Predicting Drug Responses for Vascularization of Organoids & Conventional & IO therapies Ischemic Tissues Using Mi
Slide 6: Methods to Form Spheroids/Organoids
Methods to Form Spheroids/Organoids Scaffold Scaffold-free Gel-based Multiple Non-uniform Singular Uniform Multiple Uniform Corning® Matrigel® Matrix Ultra-Low Attachment Surface Corning Spheroid Microplate Corning Elplasia® Plate Products Corning Elplasia® 12K Flask Collagen Corning Matrigel® Matri
Slide 7: Corning Elplasia technology for embryoid body formation
Corning® Elplasia® technology for embryoid body formation Average Spheroids Format (round bottom) 6-well 2,885 24-well 554 96-well 79 iPSCs 7 General - Corning (L4)
Slide 8: Bulk spheroid production: Corning Elplasia 12K Flask
Bulk spheroid production: Corning® Elplasia® 12K Flask Key Features and Benefits: • Highly reproducible bulk spheroid formation across microcavities • Compatible with brightfield and fluorescence microscopy • Scaffold-free culture • Round well-bottom geometry • Generates ~12,000 spheroids per flask
Slide 9: Corning Synthegel 3D Matrix Kits: Synthetic scaffold for iPSCs and spheroids
Corning® Synthegel™ 3D Matrix Kits: Synthetic scaffold for iPSCs and spheroids Expansion of hiPSCs in 3D (embedded & suspension) Cancer spheroids 9 General - Corning (L4)
Slide 10: Corning Synthegel 3D Matrix Kits offer ease of use across applications
Corning® Synthegel™ 3D Matrix Kits offer ease of use across applications Self-assembly Tunable Gel Defined Synthetic of peptides stiffness with X-linker Recovery of Works at room Transparent Easy to use cells and temperature matrix and handle spheroids © 2018 Corning Incorporated 10 General - Cornin
Slide 11: Corning Matribot Bioprinter: consistent dispensing of cell culture hydrogels
Corning® Matribot® Bioprinter: consistent dispensing of cell culture hydrogels Corning Matribot Bioprinter is a cost-effective unit for innovators to enter the 3D cell culture and bioprinting fields • Dispense and Print capabilities • Temperature controlled • Compact size • Intuitive programs • Mult
Slide 12: Corning Matribot Bioprinter: Dispensing for Organoid Assays
Corning® Matribot® Bioprinter: Dispensing for Organoid Assays Viable Cells Dead Cells Application note: CLS-AN-772 General - Corning (L4)
Slide 13: Summary and Resources
Summary and Resources Visit Corning 3D cell culture serves multiple disciplines Booth #601 Consistency in 3D can take on many forms: 13 General - Corning (L4)
Slide 14: Corning
CORNING
Slide 15: Predicting drug responses for conventional and IO therapies using MicroOrganoSpheres (MOS)
Predicting drug responses for conventional and IO therapies using MicroOrganoSpheres™ (MOS) Dennis Plenker, Ph.D., June 15th 2023 Dennis.Plenker@Xilis.net Proprietary and confidential – do not duplicate or distribute without written permission from Xilis, Inc.
Slide 16: Disclosures
Disclosures • Employee of Xilis Inc. • Shareholder of Xilis Inc. 2
Slide 17: Introduction
Introduction Improving predictivity in healthcare, clinical study stratification and drug discovery Marquart et al., 2018, JAMA Oncol Only ~ 10% of patients can be matched to a therapy using sequencing Only ~ 50% of all patients will respond to a therapy after sequencing-based eligibility criter
Slide 18: Only a minority of eligible patients responds to chemotherapeutic agents
Only a minority of eligible patients responds to chemotherapeutic agents Maldonado et al., 2020, Future Sci. • Median overall likelihood of response is as good as a coin flip (50%)! • Very few to no robust biomarkers available for chemotherapy response • Goal: using patient avatars for functional pr
Slide 19: World-renowned scientists and clinical researchers are on Xilis' Boards
World-renowned scientists and clinical researchers are on Xilis’ Boards, partnering with experienced leadership team and VCs to advance MOS Platform Scientific Advisory Board Clinical Advisory Board James Allison, PhD Michael Varney, PhD Scott Kopetz, MD, PhD Scott Antonia, MD, PhD Nobel Prize Winne
Slide 20: Xilis MicroOrganoSphere (MOS) technology
Xilis MicroOrganoSphereTM (MOS) technology Patient MicroOrganoSpheres (“MOS”) XilisμO Chip Day 2 Day 3 Day 4 Day 6 Proprietary IP TISSUE Rapid growth MOS Generator Drug discovery 6
Slide 21: Unique features of MicroOrganoSpheres
Unique features of MicroOrganoSpheresTM • MOS… • are mobile and can be pipetted from well to well after generation automation-friendly • are independent entity and represent a biosphere • can be used for position-specific / longitudinal imaging • can retain microenvironment for >21 days 7
Slide 22: MicroOrganoSphere Technology and applications in peer-reviewed publications
MicroOrganoSphereTM Technology and applications in peer-reviewed publications June ‘22 July ‘22 August ‘22 December ‘22 Ding et al., 2022: Wang et al, 2022: Bose et al., 2022: https://pubmed.ncbi.nlm.nih.gov/35508177 NCI MOS Feature, Online https://pubmed.ncbi.nlm.nih.gov/35985334/ https://pubmed.nc
Slide 23: Largely preserved driver mutations between tumor and MOS
Largely preserved driver mutations between tumor and MOS MOS Tissue Grey: Driver mutations present White: Driver mutations absent Venn diagrams showing fraction of shared mutations between tumors (T) and matched MOS (M) MOS Tissue MOS Tissue MOS Tissue MOS Tissue MOS Tissue MOS Tissue MOS Tissue Sen
Slide 24: Four critical MOS features: recapitulated native tumor microenvironment
Four critical MOS features unlock a tectonic shift in drug discovery and development 1 Recapitulated native tumor microenvironment TME composition preserved in MOS vs primary tumor Whole-MOS imaging reveal TME and structure scRNA-seq Flow cytometry Mesothelin, Vimentin, DAPI, CD3, CD14, CD11c, CD68
Slide 25: Four critical MOS features: recapitulated native tumor microenvironment
Four critical MOS features unlock a tectonic shift in drug discovery and development 1 Recapitulated native tumor microenvironment TME composition preserved in MOS vs primary tumor Whole-MOS imaging reveal TME and structure scRNA-seq Flow cytometry Mesothelin, Vimentin, DAPI, CD3, CD14, CD11c, CD68
Slide 26: MOS replicate endogenous immune microenvironment
1) MOS replicate endogenous immune microenvironment Flow immuno-profiling antibody staining of immune compartment Endogenous T cell tumor killing after checkpoint inhibition CD4/CD8/CD45/TIM3/PD1 12
Slide 27: MOS replicate endogenous immune microenvironment
1) MOS replicate endogenous immune microenvironment Primary lung tumor MOS Primary Primarytumor tumro MOS PD-L1 PD-1 TGFβ 13
Slide 28: Lung cancer MOS maintain endogenous immune compartment for 3 weeks
1) Lung cancer MOS maintain endogenous immune compartment for 3 weeks MOS from frozen tissue MOS immunophenotyping shows largely maintained immune compartment Red: Live MOS staining 14
Slide 29: Predict clinical response in complex IO setting
Four critical MOS features unlock a tectonic shift in drug discovery and development 2 Predict clinical response in complex IO setting Complete Response Partial Response No Response Patient-derived MOS predict patient response to IO antibody drug MOS prediction completed within 10 days of initia
Slide 30: T cell infiltration and potency assay
2) T cell infiltration and potency assay Bulk organoids – T cell can’t infiltrate MOS – T cell infiltration MOS – T cell potency 0h 24h 48h 72h Bulk Matrigel + immune cells At least four separate groups using unselected TIL have retrospectively evaluated the interferon release assay but found that i
Slide 31: Prospective clinical study demonstrating clinical predictability of MOS IO assay
2) Prospective clinical study demonstrating clinical predictability of MOS IO assay for checkpoint inhibitors and combos Xilis’ clinical IO assay completed within 10 days of initial diagnosis predicted ensuing patients’ response (20 patients to date) Example #1: Example #2: Hepatocarcinoma patient
Slide 32: Versatile across therapeutic modalities & tumor types
Four critical MOS features unlock a tectonic shift in drug discovery and development 3 Versatile across therapeutic modalities & tumor types chemo targeted IO Cell Therapy T-cell killing TIL-mediated MOS killing (green) SN38 Daratumumab Nivo TIL 18
Slide 33: MOS can help overcome low cell number, heterogeneity, and low throughput
3) MOS can help to overcome low cell number, heterogeneity, and low-throughput of patient-derived models Whole-well drug response quantification ML-driven MOS drug response quantification (normalization) ATP-based Brightfield FL signal Increasing dose IC50 19
Slide 34: Scalable and automatable for drug discovery
Four critical MOS features unlock a tectonic shift in drug discovery and development 4 Scalable and automatable for drug discovery Drug response across patient MOS Scaled MOS automation enabled a study that tested 215 drugs from 1,000 past CRC trials and identified winners and losers in 9 days. 20
Slide 35: Automated MOS screening and AI analytical pipeline: 1000 colon cancer trials in 9 days
4) Xilis’ automated MOS screening and AI analytical pipeline offer the first high- throughput patient-derived model platform – 1000 colon cancer clinical trials in 9 days with MOS patient avatars Screening Metrics 215 Compounds (8 concentrations) ~40,000 Conditions screened / day 5,806,080 Images 50
Slide 36: Bose et al, Cancer Cell, 2022
Bose et al, Cancer Cell, 2022 22
Slide 37: Outlook
Dennis.Plenker@Xilis.net Outlook Registered multi-center clinical study show MOS predict SoC chemo – CRC, breast Predicted by MOS: Ding et al, Cell Stem Cell, 2022 MOS predict refractory multiple myeloma patient clinical response • Utility of MOS for Multiple Myeloma for clinical correlation • Predi
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Slide 39: Applications of Human Pluripotent Stem Cell-derived Vascular Cells
Applications of Human Pluripotent Stem Cell- derive Vascular Cells Ping Zhou Associate Adjunct Professor Director, Stem Cell Core Stem Cell Program University of California Davis
Slide 40: The authors declare no potential conflicts of interest
The authors declare no potential conflicts of interest.
Slide 41: Stem Cell Cores At University of California Davis
Stem Cell Cores At University of California Davis Pluripotent Stem Cells (PSCs) Embryonic Stem Cells (ESCs) Induced Pluripotent Stem Cells (iPSCs) Multipotent Stem Cells Mesenchymal stem cells (MSCs) Hematopoietic Stem cells (HSCs) Primary Cells • Generate, expand and store human pluripotent stem ce
Slide 42: Vascular Disease
Vascular Disease 8 million people in the United States Limb ischemia
Slide 43: Revascularization of the ischemic tissue is needed to treat vascular disease
Revascularization of the ischemic tissue is needed to treat vascular disease. Transplantation of vascular cells may accelerate revascularization.
Slide 44: Human pluripotent stem cells can be an unlimited source of vascular cells
Human pluripotent stem cells (PSCs) can be an unlimited source of vascular cells for therapy.
Slide 45: Vascular cell types
Pericyte Red blood ‘@\ cell in lumen Basement ff membrane Endothelial cell Capillaries Arteriole Venule Artery Endothelial cells Smooth muscle cells Smooth muscle cells
Slide 46: Differentiation of Endothelial Cells from Human PSCs
Differentiation of Endothelial Cells from Human PSCs VEGF BMP4 FGF2 FGF2 EGF GSK3βi VEGF IGF1 2 days 2 days 4-6 days iPSC/hESCs Mesoderm Vascular Endothelial Progenitors Cells
Slide 47: hiPSC-ECs and hESC-ECs CD31 staining
Phase CD31 hiPSC-ECs |; hESC-ECs |
Slide 48: hESC-ECs and hiPSC-ECs flow cytometry
hESC-ECs hiPSC-ECs 10*= 10? “H1 H2 -H1 H2 . ~ 102 0.1% 78.3% sed rae + 402- 0.2% 83.1% : pipe = 3 = , F by © 10! : © 10! oe oH HAR > H3 Hae 10 2.8% 18.8% 10 ee 14.3% 700 ~~ 401 ~~ 402 “403 70° 01 ~~" 402 “403 CD31 CD31
Slide 49: The Human PSC-derived Endothelial Cells Formed Tubes in Matrigel
The Human PSC-derived Endothelial Cells Formed Tubes in Matrigel In vitro In vivo
Slide 50: Mouse Model of Ischemia
Mouse Model of Ischemia FITC-Dextran Tail Vein Injection 1 day 2 months Immune Deficient Femoral artery Intramuscular NSG Mice ligation to create injection limb ischemia of PSC-ECs
Slide 51: iPSC-derived Endothelial Cells Generated Functional Blood Vessels in Ischemic Mouse Model
IPSC-derived Endothelial Cells Generated Functional Blood Vessels in Ischemic Mouse Model FITC-Dextran Human CD31 Merge
Slide 52: Differentiation of Vascular Smooth Muscle Cells/Pericytes Progenitors from Human iPSCs
Differentiation of Vascular Smooth Muscle Cells/Pericytes Progenitors (VSMPs) from Human IPSCs Endothelial Cells (ECs) Vascular hiPSCs Progenitors Vascular Smooth Muscle/ Pericyte Progenitors (VSMPs)
Slide 53: Human PSC-derived Vascular Cells Formed Tubes on Matrigel
Human PSC-derived Vascular Cells Formed Tubes on Matrigel Green: endothelial cells Red: smooth muscle cell/pericyte progenitors
Slide 54: Can we use human iPSC-derived ECs to deliver therapeutic proteins?
Can we use human iPSC-derived ECs to deliver therapeutic proteins?
Slide 55: Hemophilia A (HA)
Hemophilia A (HA) • Spontaneous or prolonged bleeding leads to damage in joint, nerve and tissue. • Bruise easily and delayed wound healing • HA affects 1 per 5000 males. • HA is caused by mutations in the coagulation factor VIII gene. • Factor VIII is produced primarily by endothelial cells.
Slide 56: FVIII Replacement Therapy
FVIII Replacement Therapy 2-3 times of infusions FVIII neutralizing Life-long Cost per week for life-long antibodies $300,000/year
Slide 57: HA Patient iPSCs
HA Patient iPSCs F8 Expressing iPSCs Endothelial Cells
Slide 58: ECs Derived from Hemophilia A Patients' iPSCs Did Not Express F8
Endothelial Cells (ECs) Derived from Hemophilia A Patients’ IPSCs Did Not Express F8
Slide 59: Expression of the F8, GFP and Luciferase Gene in HA-iPSC-derived ECs Using Lentiviruses
Expression of the F8, GFP and Luciferase Gene in HA- iPSC-derived ECs Using Lentiviruses Lentivieuse vectors: pCCLc-MNDU3-F8-PGK-neo pCCLc-MNDU3-LUC-PGK-GFP
Slide 60: HA-iPSC-derived ECs Engrafted in NSG Mice
HA-iPSC-derived ECs Engrafted in NSG Mice
Slide 61: HA-iPSC-derived ECs Overexpressing F8 Produced Therapeutic Levels of FVIII in HA Mice
HA-iPSC-derived Endothelial Cells Overexpressing F8 Produced Therapeutic Levels of FVIII in HA Mice 2 weeks Analysis
Slide 62: Generation of Liver Organoids from Human PSCs
Generation of Liver Organoids from Human PSCs PSCs Differentiate in vitro Hepatic Cells Endothelial Cells Co-culture on solidified Matrigel in vitro Liver Organoids
Slide 63: Applications of PSC-derived Liver Organoids
Applications of PSC-derived Liver Organoids Disease Modeling Cell Therapies Drug Screening Toxicity Test
Slide 64: Organoids from ESCs and Organoids from iPSCs
Organoids from ESCs _ Organoids from iPSCs Wy HP+EP+MSC
Slide 65: Liver Organoids Derived from Human PSCs
Liver Organoids Derived from Human PSCs Hepatic cells (AFP) Endothelial Cells (CD31)
Slide 66: Liver Organoids Derived from Human PSCs
Liver Organoids Derived from Human PSCs iPSC-derived hESC-derived Two months after transplantation
Slide 67: Liver Organoids Expressed Liver Markers In Vivo
Liver Organoids Expressed Liver Markers In Vivo
Slide 68: Summary
Summary • Vascular cells could be derived from human PSCs and capable of generating functional blood vessels in ischemic tissues. • Patient iPSC-derived endothelial cells could be used to deliver therapeutic factor VIII proteins for the treatment of hemophilia A. • Liver organoids could be generated
Slide 69: Acknowledgement
Acknowledgement Zhou Lab Collaborators Nataly L Magner Dr. Jan Nolta Dane Coleal-Bergum Dr. Aijun Wang Aja Harding Dr. Ben Waldau Elizabeth Cortez-Toledo Melanie Rose Emmanuel Agu Alicia Hyllen Kelsey Conroy Financial Support Amy Kim UC Davis Innovative Development Award Dickerson’s Catalyst Fund NI
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