Tengion Corporate Overview

BIOACTIVE RENAL CELLS
AUGMENT KIDNEY FUNCTION IN A RODENT
MODEL OF CHRONIC KIDNEY DISEASE
Rusty Kelley, PhD
Regenerative Medicine & Biology
Tengion Labs
May 24, 2010
ISCT Annual Meeting, Philadelphia PA
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Product Pipeline – May 2010
Optimization
Preclinical
IND
Neo-Bladder Augment
Spina Bifida
Spinal Cord Injured
Urge Incontinence (laparascopic)
Neo-Urinary Conduit
Bladder Cancer
Neo-Bladder Replacement
Bladder Cancer
Neo-Kidney Augment
Advanced CKD
Neo-GI Augment
Esophagectomy
Neo-Vessel Replacement
Vascular Access Graft
Peripheral Artery Bypass
Coronary Artery Bypass
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Phase I
Phase II
Phase III
Chronic Renal Failure is a leading cause of death worldwide
New treatment modalities are needed
http://www.nationmaster.com/red/pie/mor_chr_ren_fai-mortality-chronic-renal-failure
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Goal: Identify bioactive cellular components for
Neo-Kidney Augment™ (NKA) prototypes
Through in vitro characterization and in vivo testing
Strategic Approach:
1) Generate testable array of
‘kidney components’ based
on native tissue composition
2) Design admixture
experiments based on
functional component
characteristics
3) Test admixture arrays in vivo
in small animal model of
terminal, progressive CKD
2-step
5/6 Nx
(Rodent)
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*patent filed: 2009
Isolation and propagation of endocrine, tubular, and
glomerular cells has been described from healthy rodent
kidney tissue
Work completed via Sponsored Research Agreement
with Wake Forest Institute of Regenerative Medicine
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Heterogeneous cultures of kidney cells (UNFX)
Fractionated based on differences in buoyant density
CKD or non-CKD
FRACTIONS
Enzymatic Digestion
Cell Culture/Expansion
Gradient Separation
UNFX
a heterogeneous mixture of renal cells isolated cortical and medullary zones
B1
distal tubular and collecting duct cells with trace amounts of other cell types
B2
renal tubular epithelial cells with distal tubule and collecting duct cells present.
Endocrine, glomerular and vascular cells are present in trace quantities
B3
proximal tubular cells, with fewer distal tubule, collecting duct, endocrine,
vascular, glomerular, and progenitor-like cells
B4
vascular, endocrine, glomerular with fewer tubular and progenitor-like cells
B5
very small cells with low viability
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In vitro characterization of subfractions
Identified unique properties of B2 and B4
• Subfraction B2 is enriched for tubular cells
• Subfraction B4 is enriched for vascular, glomerular, and oxygenregulated erythropoietin (EPO)-producing cells
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Microarray gene expression analysis of Subfractions
Validated rat expression and confirmed human translation by qrtpcr
Validation of microarray by qRTPCR
David* Annotated Functional Group: Blood Vessel Development
Target Gene
CDH5
-KDR
KDR
PLAT
ANGPT2
Sample
Rat RQ
Human
CKD RQ
Human
Non-CKD RQ
B2
0.742
1.052
1.387
B4
8.065
6.205
2.340
B2
0.708
0.607
0.233
B4
10.348
20.637
6.344
B2
1.008
1.224
0.430
B4
3.088
4.266
0.430
B2
0.737
0.872
0.812
B4
6.697
14.115
13.446
B4-specific gene expression patterns were validated for
the following David* Annotated Functional Groups:
• Blood vessel development
• Extracellular matrix
• Regeneration
• Localization of the cell
• Developmental processes
*http://david.abcc.ncifcrf.gov
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In vivo testing of subfractions & admixtures
In a 2-step 5/6-nephrectomized (NX) rodent model
4-7 WEEKS
2-step 5/6 Nephrectomy
Day 0
Day 7
Poles of Left
Kidney removed
(2/6th)
Right Kidney
Removed
(3/6th)
TREATMENT
Establishment
of Disease State
Entry Criteria
sCREAT & BUN
monitored weekly
↑ sCREAT ≥200%
↑ BUN ≥150%
≥2 consecutive weeks
CONTROLS TEST
Model Generation
12-24 WEEKS
RANDOMIZATION
1 WEEK
Intrarenal delivery of cells in diluent
No Surgery & Sham Nx
No treatment
FOLLOW-UP
• serum chemistry
• urinalysis
• hematology
• survival
• weight gain
• kidney weight
• eGFR
• Blood Pressure
• histology
• Animals were treated after disease state established
• Series 1: B2 and B4 tested independently in comparison to UNFX
• Series 2: Subfraction admixtures tested in comparison to B2
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Selected subpopulations enhanced systemic health
Survival and weight gain
• B2 > B4 > UNFX for survival and weight gain
• Treatment with B2 à 100% survival at 6 month endpoint
Series 1
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Selected subpopulations enhanced renal function
Filtration, functional renal mass, and erythropoiesis
B2 > B4 > UNFX
Series 1
• Stabilized serum creatinine
• Enhanced functional renal mass
• Supported erythropoiesis
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Summary of Results
Series 1: Comparison of UNFX, B2, & B4
• When data are considered across multiple parameters
tested, the B2 cellular prototype provides significant and
reproducible therapeutic benefits compared to UNFX and B4
§
§
§
Survival
Filtration
Erythropoiesis
• The B4 cellular prototype provides a mild survival benefit as
well as supports erythropoiesis and facilitates glomerular
repair.
• Thus, the B2 subpopulation served as comparison for
Series 2 – tests of subpopulation admixtures
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Subpopulation admixture B2+B4
Provided superior therapeutic outcomes in vivo
NS = No Survival
Series 2
B2+B4
• Improved eGFR
• Stabilized serum Creatinine & BUN
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Therapeutic effects of B2+B4 were confirmed histologically
And compared to NX and an admixture of B1+B5
• Remodeling and regeneration in tubular and glomerular compartments
•
Modulation of glomerular and tubulo-interstitial fibrosis
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Summary of Results
Series 2: Testing of subpopulation admixtures
The B2+B4 admixture outperformed other cell and cell-cell
combinations tested on multiple systemic and histological
parameters:
§ Weight gain
§ Survival (100%)
§ Lowering serum creatinine and BUN
§ Raising estimated Glomerular Filtration Rate (eGFR)
§ Lowering systemic blood pressure
§ Reducing glomerular injury
§ Reducing tubular injury
§ Stimulating tubulogenesis
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Conclusions
§
Specific subpopulations of renal cells are more efficacious and reproducible
in slowing the progression of CKD than unfractionated (UNFX)
§
Tubular cell-enriched subpopulation B2 improves functional renal mass
significantly and leads to organism-level benefits, including survival
§
While B4 is less therapeutic on its own, an admixture of B2+B4 enhances the
therapeutic outcome of B2, providing superior
§ Renal filtration
§ Tubular regeneration
§ Glomerular repair
§
B2-based admixtures (e.g., B2+B4) provide a regenerative stimulus for the
treatment of renal insufficiency
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