Organovo Brochure - The Company's Information
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Transcript of Organovo Brochure - The Company's Information
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7/27/2019 Organovo Brochure - The Company's Information
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Copyright 2013, Organovo Holdings, Inc. This report is solely for the use of intended audience. No part
of it may be circulated, quoted, or reproduced for distribution outside the organization without prior writtenapproval from Organovo Holdings, Inc.
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SAFE HARBOR STATEMENT
1
OTCQX:ONVO
Copyright 2013 Organovo, Inc.
This presentation may contain forward-looking statements that involve significant risks
and uncertainties. All statements other than statements of historical fact are forward-
looking statements, including statements regarding the Companys prospectiveperformance, opportunities and business outlook. Any forward-looking statements
contained herein are based on current expectations, but are subject to a number of risks
and uncertainties. The factors that could cause the Companys actual future results to
differ materially from current expectations include, but are not limited to, risks and
uncertainties relating to the Company's ability to develop, market and sell products
based on its technology; the expected benefits and efficacy of the Companys productsand technology; the availability of substantial additional funding for the Company to
continue its operations and to conduct research and development, clinical studies and
future product commercialization; and the Company's business, research, product
development, regulatory approval, marketing and distribution plans and strategies.These and other factors are identified and described in more detail in our filings with the
SEC, including our annual report for the period ended December 31, 2012 on Form 10-
K and our current reports filed on Form 8-K. These forward-looking statements are
made as of the date of this presentation, and we do not undertake an obligation toupdate these forward-looking statements after such date.
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Current in vitro cell-based modelsare often poor predictors ofin vivo outcomes
2
Attrition rates for experimental andapproved drugs remains high
Cardio- and hepatotoxicity are theprimary causes for late-stage failures
and post-market withdrawals
More robust human models of theseorgan systems are needed
Copyright 2013 Organovo Holdings, Inc.
CardiovascularToxicity
40%
Other33%
Hepatotoxicity27%
From: EvaluatePharma; CDER; Tufts Center for Drug Discovery
Drugs withdrawn from global market due to toxicity
1990-2010 (n=39)
Hepatoxicity;
26%
CVToxicity;
33%
Other;41%
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Organovos NovoGen BioprinterTM platform
was utilized to generate a 3D human liver system
3
Design concept for 3D human liver
focused on the following key features:
True three-dimensionality, reaching atleast 250 microns in the smallestdimension
Incorporation of multiple cell types withspatially-controlled placement in x, y,
and z axes
Demonstration of both histologic andfunctional features of liver
Copyright 2013 Organovo Holdings, Inc.
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Materials and Methodsutilized in these studies
4
Cell Culture. All cells utilized in these experiments were sourced from commercial vendors and culturedaccording to manufacturers recommended protocols. Cryo-preserved primary human hepatocytes were purified
by Percoll gradient centrifugation prior to use in bioprinted constructs. iPSC-derived hepatocyte-like cells (iCells)were generously provided by Cellular Dynamics, Inc.
Bio-ink Preparation. Bio-ink was prepared based on protocols and techniques described in Norotte et al.,(Biomaterials 30:5910-5917). For preparation of hydrogels containing cells, NovoGel 2.0 (Organovo, Inc.) wasprepared according to manufacturers instructions and non-parenchymal cell populations were incorporated prior
to bioprinting.
Bioprinting.All tissues were fabricated directly into standard tissue culture plates (Corning Transwell) usingstandard Organovo bioprinting protocols and the NovoGen MMX BioprinterTM or modifications thereof.
Secreted protein detection. Spent media was analyzed by commercially available ELISA kits for the followingliver proteins: albumin, fibrinogen, and transferrin. Cholesterol biosynthesis was quantified fluorimetrically
(Cayman Biochemical).
CYP 450 analysis. CYP1A2 and CYP3A4 activities were assessed with the Pro-GloTM CYP450 Assay system(Promega). Bioprinted liver tissues were challenged with either verapamil (10M) or dexamethasone (10M) to
stimulate CYP1A2 or CYP3A4 activity. Fold induction was calculated as the increase in activity of the treatedsamples relative to the non-treated control samples.
Histologic analysis. Tissues were fixed in 10% buffered formalin, paraffin-embedded, and subjected tostandard histochemical analyses. In some experiments, tissues were snap frozen upon harvest and
cryosectioned prior to histologic or immunohistologic analysis.
Copyright 2013 Organovo Holdings, Inc.
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3D liver tissues with relevant architectural features
were fabricated successfully with the NovoGen Bioprinter
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Single- or Multi-unit bioprinted liver tissues were constructed from hepatocytes or
hepatocyte-like cells and non-parenchymal cells, and were characterized by a tissue-
like cellular density and tight intercellular junctions.
Copyright 2013 Organovo Holdings, Inc.
Single-Unit Geometry Multi-Unit Geometry Multi-Unit Geometry
H&E (20x) E-Cadherin (20x) Multi-Unit Tissue(post-fusion)
100 m100 m
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Bioprinted 3D human liver tissuesare metabolically active with CYP450 induction
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These liver tissues remained stable for
135 hours and retained key liverfunctions, including inducible CYP1A2
and CYP3A4 activity.
Copyright 2013 Organovo Holdings, Inc.
Single Unit / 39 hrs Single Unit / 135 hrs
Constructed w/ 1 hepatocytes, hepatic stellate cells, and endothelial cells
135Hrs
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Bioprinted 3D human liver tissuessynthesize cholesterol
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Bioprinted liver tissues possess the ability
to synthesize cholesterol, a hallmarkliver-associated function. Cholesterol
biosynthesis is retained and evenincreases over time in culture.
Copyright 2013 Organovo Holdings, Inc.
Single-Unit Geometry / 48 hrs
Constructed with HepaRG cells, hepatic
stellate cells, and endothelial cells
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The NovoGen Bioprinter builds complex 3D tissuewith precise deposition of distinct cell populations
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Bioprinted human liver tissues are characterized by cell-specific
compartments that enable reproduction of native tissue-like architecturalpatterns and the establishment ofcontrolled cell-cell interfaces.
Copyright 2013 Organovo Holdings, Inc.
2.5x (light microscope image)
Hepatic stellate cells (CFSE)
Hepatocytes (DAPI)
Endothelial cells (CTMB)100 m
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3D human liver tissues outperform 2D cultureswith respect to liver-specific protein production
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Bioprinted liver constructs, generated from
iPSC-derived hepatocyte-like cells (iCells;
Cellular Dynamics, Inc.), are well organized,develop microvascular networks over time,
and produce significantly more albumin
per cell than matched 2D controls.
Copyright 2013 Organovo Holdings, Inc.
H&E / 20x100 m
CD31 / 20x100 m
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3D human liver tissues were fabricated using an automated bioprintingplatform to yield distinct geometries which include both simple and repeatingfunctional units
Constructs contained heterogenous cellular inputs including hepatocytes,hepatic stellate cells and endothelial cells
Bioprinted constructs were organized, dense and viable, exhibitingmicroarchitecture in a manner approximating in vivo tissues
Sustained production of a number of critical functional endpoints has beendemonstrated
Albumin, fibrinogen, transferrin, cholesterol, CYP activity The platform enabled generation of functional constructs from a variety of cell
sources, including primary hepatocytes and stem/progenitorcell-derived
hepatocyte-like cells (HepaRG, iCells) Per-cell production of a liver-specific protein Albumin was significantly
enhanced in 3D liver constructs vs. matched 2D cultures
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Summary of Experimental Observations
Copyright 2013 Organovo Holdings, Inc.
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Conclusions
11 Copyright 2013 Organovo Holdings, Inc.
H&E / 20x
Bioprinting enabled highly reproducible fabrication of architecturally andcompositionally defined 3D tissues into standard tissue culture formats
Bioprinted 3D liver tissues exhibited several key unique features that remainedstable over time
Tissue-like cellular density with high viability and well-organized microarchitecture(microvasculature, tight junctions)
Cell type-specific compartmentalization, with establishment and retention of user-defined spatial localization of parenchymal and non-parenchymal cells
Multi-layered architecture ranging from 250-500 microns in thickness 3D liver tissues possessed critical liver functions, including albumin production,
cholesterol biosynthesis, fibrinogen and transferrin production, and inducible
CYP1A2 and CYP3A4 activities
Per-cell production of Albumin by 3D bioprinted tissues was 5-9x greater thanmatched 2D controls, suggesting superior functional performance in 3D