organovo brochure - the company's information

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

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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

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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

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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



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Materials and Methodsutilized in these studies

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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.



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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.


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.


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.


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.


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.


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



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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

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