tristar technology group corporate presentation
DESCRIPTION
TRANSCRIPT
TriStarTechnology Group
9700 Great Seneca Highway, suite 401Rockville, MD 20850(E) [email protected](P) 301-792-633 (W) www.tristargroup.us
Sample Acquisition With Annotated Clinical Information
A Critical Success Factor In Biomarker Validation
the need for targeted therapeutics with companion diagnostics
Development of targeted therapeutics requires testing in targeted populations matched to a drug’s mechanism of action
Evaluation of Trastuzamab in “all comer” breast cancer patients (25% HER+, 75% HER2-) would not have shown significant benefit in clinical trials
Early proof of concept in the right patient population is crucial
Potentially shorter time to market
An emerging unmet need in oncology drug development today is service providers that offer both access to well-annotated specimens and sophisticated molecular analytical capabilities
Rockville, MD
Rome & Catania, Italy
Hamburg, Germany
Madrid, Spain
TMA Repository
TMA Repository TMA Repository & Contract Research
Array ManufacturingContract Research
Cancer Stem Cell Research
tristar providesAccess to 2.5 million archived samples & clinical data
Access to patients (prospective collection projects)
Fit-for-purpose analytical platforms & services (IHC, FISH, qRT-PCR etc.)
Collaboration for Solid tissue biomarker development
ethical considerations
Informed Donor Consent
IRB/EC Approval
Fully Anonymized
Compliant with Current International & EU Regulations
Blocks That Are in Excess of Diagnostic Sample Only
Team of 17 Pathologists & 5 Oncologists for Clinical Data Review
product groupsArchived Human Tissue Repository
>2.5 million samples (FFPE & Frozen). 70% Oncology, 30% CNS, GI etc.
High-Density Tissue Micro Arrays >100,000 donor samples with outcome data
Outcome Data Treatment, Response rates, disease –free survival (DFS), overall survival (OS)
Molecular DataER/PR/HER2, p53, BRAF, KRAS, EGFR, PIK3CA etc.
Cancer Stem Cell ArraysLysates & RNA
Blocks & Large sectionsWith matching RNA, DNA
Protein ExpressionIHC (Antibody protocol development, automated or manual staining, reading & interpretation)
Gene ExpressionRT-PCR
Gene copy numberFISH/CISH
Cross-Reactivity Screening in Normal Tissue(GLP)
Large-Scale Analysis of Prognostic markers (500-3500 donor samples)(500-3500 donor samples)
Gene sequencingDNA sequencing
our services
quality controlSamples are fixed/frozen within 2 – 10 minutes of Excision
OCT embedded sample
Snap frozen sample
Formalin fixed sample
quality control10% Buffered formalin, 10-12 hrs. fixation timeMorphology (H&E) & IHC Markers for immunogenicity
RNA & DNA Quality (Agilent 2100 Bioanalyzer)
RIN can be checked & provided upon request
Primary Tumors Matched Mets Approximate number
BreastNodal
DistantBone
200020
200
CRC NodalLiver
2000150
Prostate NodalBone
500300
Lung (NSCLC) NodalBone
300100
Pancreatic Nodal 100
Head & Neck Nodal/Soft tissue 100
Gastric Nodal, liver etc 200
Melanoma Nodal 50
primary tumors with matched mets
tumor type data approximate number
Breast5 yr survival
10 yr. survivalHerceptin
5000300
400 (responders & non-responders)
CRC3-5 yr survival
Bevacizumab, Cetuximab4000
500 (responders & non-responders)
Prostate,Breast, CRC, Ovarian
10 yr survivalSOC Chemotherapy
50001500 (responders & non-responders)
Lung (NSCLC)3-5 yr survival
Docetaxel, Gemcitabine2000400
Pancreatic Survival 350
Head & Neck Treatment/survival 200
Gastric Survival 250
NHL Survival 200
Ovarian 3-5 yr. survival 300
Bladder Survival 500
samples with outcome data
Formalin Fixed Paraffin Embedded
Frozen OCT Embedded
tissue microarrays
Morphology
RNA/Protein/DNA
DNA
tissue microarrays to study tumor heterogeneity
The whole tumor is sectioned into 8-10
constituent blocks The exact localization of each
block is recorded
Cores are taken from each constituent tumor block and transferred to a TMA.
Tumor Type Primary tumors
Blocks per tumor
Matched Nodal Mets
Blocks per met
Total number of TMA Cores
NSCLC 146 8 66 4 1432
Breast 147 8 32 4 1304
CRC 140 8 42 4 1288
Prostate 190 10 - - 1900
Bladder 147 8 - - 1176
tissue microarrays to study tumor heterogeneity
Allows for an overview of the whole tumorAn optimal way to measure intratumoral heterogeneity
Heterogeneity found in 7/13 (54%) EGFR amplified NSCLC
EGFR FISH Result
amplification
normal
polysomy
n.a.
Case
#1
#2
#3
#4
#5
#6
#7
Different areasof the primary cancer
Different matched lymph node metastases
EGFR amplification is often heterogeneous in lung cancer
PTEN + ERG
PTEN only
ERG only
PTEN deletions are late events developing preferentially in ERG
positive prostate cancers
heterogeneity TMA: co-analysis of ERG and PTEN in prostate cancer
35 ERG+PTEN10 PTEN only4 ERG only PTEN linked to ERG
P<p<0.0001
31 tumors PTEN+ERG21 ERG precedes PTEN0 PTEN precedes ERG earlier
Frequency of PTEN deletion is strongly linked to prostate cancer progression (n >2200 donor samples)
p<0.0001
PIN (n=29)
BPH (n=20)
pT2 (n=1085)
pT3a (n=360)
pT3b (n=227)
pT4 (n=24)
HR (n=54)
0.0
5.0
10.0
15.0
20.0
25.0
30.0
35.0
40.0
45.0
50.0
PTEN homozygous
PTEN hemizygous
fracti
on o
f tu
mors
(%
)
prostate cancer progression & prognosis analysis
tissue micro arrays to study tumor heterogeneity
The level of heterogeneity of therapy target genes may be relevant for diagnosis and response
HER2 is homogenous in breast cancer but heterogeneous in colon cancer
Tumor heterogeneity is clinically important and can be optimally addressed by heterogeneity TMAs
What normal tissues do express target?
How frequent is expression in human cancer?
Option 1:Review the literature
Specific cancer subtypes or biological properties?-prognostic relevance
Option 2:Perform
own studies
Most oncology drugs in development are expected to be active only in sub-sets of patients
molecular epidemiology
Prognosis TMA-Based Target Evaluation Strategy (IHC)
Multi-Tumor Tissue Array3,500 donor samples
All Cancer Types
Normal Tissue Array600 donor samples
532 Cell Types
Tumor Cell Line Array140 Cell Lines
Including NCI 60
Cancer – Specific Prognosis TMA Analaysis
Breast Cancer(2,000 donors)
Prostate Cancer(3,000 donors)
Lung Cancer(1,400 donors)
Bladder Cancer(1,100 donors)
Pancreatic Cancer(300 donors)
Colon Cancer(1,400 donors)
Ovarian Cancer(200 donors)
NHL(200 donors)
Relationship of Molecular Target to Prognosis, Histological Sub-type, Response to Treatment etc
TriStar: a new dimension in tissue biomarker analysis
√ Expression in cancer types (including niche cancers)√ Complete normal tissue expression information√ Cell lines identified for functional studies/drug screening
Skin: Squamous Cell Carcinoma, Basal Cell Carcinoma, Merkel Cell Carcinoma. Uterine Corpus: Endometrioid Adenocarcinoma, Serous. Parathyroid Gland: Adenoma, Carcinoma. Mammary Gland: Intraductal Carcinoma, Lobular Carcinoma In Situ, Invasive Ductal Carcinoma, Invasiv Lobular Carcinoma, Mucinous Carcinoma, Papillary Carcinoma, Tubular Carcinoma. Kidney: Clear Cell Type, Papillary Type, Chromophobe Cell Type. Urinary Bladder: Non-Invasive Papillary Tumor (Pta), Transitional Cell Carcinoma, Squamous Cell Carcinoma, Adenocarcinoma, Small Cell Carcinoma. Salivary Glands: Mixed Tumor, Adenolymphoma, Adenoma, Mucoepidermoid Carcinoma, Acinic Cell Carcinoma, Adenocarcinoma, Adenoid Cystic Carcinoma. Esophagus: Squamous Cell Carcinoma, Adenocarcinoma. Stomach: Adenocarcinoma Diffuse Type, : Adenocarcinoma Intestinal Type. Adrenal Gland: Adrenal Cortical Adenoma, Adrenal Cortical Carcinoma, Pheochromocytoma. Pancreas: Adenocarcinoma, Adenoma. Mediastinum: Thymoma. Small Intestine: Adenocarcinoma, Carcinoid. Large Intestine: Adenoma, Adenocarcinoma. Appendix: Adenocarcinoma, Carcinoid. Anal: Small Cell Carcinoma. Prostate: Prostatic Adenocarcinoma Untreated, Hormone Refractory Adenocarcinoma Adenocarcinoma, Clear Cell Adenocarcinoma, Atypical Hyperplasia. Cervix: Squamous Cell Carcinoma, Adenocarcinoma. Vagina: Squamous Cell Carcinoma, Adenocarcinoma. Vulva: Squamous Cell Carcinoma. Thyroid Gland: Follicular Carcinoma, Papillary Carcinoma, Anaplastic Carcinoma, Medullary Carcinoma, Adenoma. Lung: Squamous Cell Carcinoma, Adenocarcinoma, Undifferentiated Large Cell Carcinoma, Small Cell Carcinoma, Carcinoid. Testis: Seminoma, Teratoma, Embryonal Carcinoma, Choriocarcinoma, Yolk-Sac-Tumor, Teratocarcinoma. Ovary: Serous Carcinoma, Mucinous Carcinoma, Endometrioid Carcinoma, Brenner Tumor, Germ Cell Tumors. Liver: Hepatocellular Carcinoma, Cholangiocarcinoma. Fibrohistiocytic: Fibrosarcoma, Benign Histiocytoma, Dermatofibrosarcoma Protuberans, Atypical Fibroxanthoma, Malignant Fibrous Hiostiocytoma Lipomatous: Lipoma, Lioposarcoma. Smooth Muscle: Leiomyoma, Leiomyosarcoma, Leiomyoblastoma. Skletal Muscle: Rhabdomyoma, Rhabdomyosarcoma. Blood And Lymph Vessels: Angioma, Epitheloid Hemangioma, Hemangioendothelioma, Angiosarcoma, Kaposi Sarcoma. Perivascular: Glomus Tumor, Hemangiopericytoma. Synovial: Benign Giant Cell Tumor Of Tendon Sheath, Synovial Sarcoma. Mesothelial: Solitary Fibrous Tumor Of Pleura And Peritoneum, Adenomatoidtumor, Malignes Mesothelioma. Neural: Neurofibroma, Neurinoma. Granular Cell Tumor, Malignant Peripheral Nerve Sheath Tumor. Clear Cell Sarcoma. Paraganglioma, Ganglioneuroma. Pnet: Ganglioneuroblastoma, Neuroblastoma, Neuoepithelioma, Extraskelettal Ewings-Sarcoma. Malignant Mesenchymoma. Alveolar Soft Part Sarcoma. Epitheloid Sarcoma. Osseous: Osteoidosteoma, Osteoblastoma, Osteosarcoma. Chondrous: Chondroblastom, Chondrom, Chondrosarcoma, Chordomas. Ewing Sarcoma. Giant Cell Tumor Of The Bone. Brain: Astrocytoma, Glioblastoma Multiforme, Oligodendroglioma, Ependymoma, Medulloblastoma, Medulloepithelioma, Craniopharyngeoma, Esthesioneuroblastoma, Retinoblastoma. Nevus Naevocellularis, Malignant Melanoma, Gastrointestinal Stromatumor, Endometrioid Stromal Sarcoma, Mixed Malignent Mesodermal Tumor, Aml, Cml, Cll, Immunocytic Lymphoma, Plasmocytoma, Centrocytic Lymphoma, Centroblastic Centrocytic Lymphoma, Centroblastic Lymphoma, Immunoblastic Lymphoma, Burkitt Lymphoma, T-Cell Lymphoma Low Grade, T-Cell Lymphoma High Grade, M Hodgkin Lymphocytic Depletion, M Hodgkin Mixed Cell Type, M Hodgkin Nodular Sclerosing etc.
multi tumor analysis including less prevalent tumor types
All tumors & sub-types are stained. Customer can selectand pay for data on specific tumors of interest
Ovarian cancer
Gall bladder cancer
Endometrial cancer
Pancreatic cancer
Urinary bladder cancer
0.0 2.0 4.0 6.0 8.0 10.0 12.0 14.0 16.0 18.0
Fraction of HER2-amplified samples (%)
Tapia et al., Modern Pathology, 20(2), 192–198 (2007)
IHC FISH
HER2 Expression and Amplification in Human Cancers
76 tissue types, 532 cell types, 8 donors each
Mesenchymal tissues: aorta/intima, aorta/media, heart (left ventricle), sceletal muscle, sceletal muscle/tongue, myometrium, appendix (muscular wall), esophagus (muscular wall), stomach (muscular wall), ileum (muscular wall), colon descendens (muscular wall), kidney pelvis (muscular wall), urinary bladder (muscular wall), penis (glans/corpus spongiosum), ovary (stroma), fat tissue (white),
Surfaces: skin (surface), skin (hairs, sebaceous glands), lip (epithelium), oral cavity, tonsil (surface epithelium), anal canal (skin), anal canal (transition epithelium), exocervix, esophagus, kidney pelvis, urinary bladder, amnion/chorion, stomach (antrum), stomach (fundus and corpus), small intestine, duodenum, small intestine, ileum, appendix, colon descendens, rectum, gallbladder, bronchus, paranasal sinus. Solid organs: lymph node, spleen, thymus, tonsil, liver, pancreas, parotid gland, submandibullary gland, sublingual gland, lip (small salivary gland), duodenum (Brunner gland), kidney cortex, kidney medulla, prostate, seminal vesicle, epididymis, testis, lung (parenchyma), lung (bronchial glands), breast, endocervix, endometrium (proliferation), endometrium (secretion), fallopian tube, endometrium (early decidua), ovary (stroma), ovary (corpus luteum), ovary (follicular cyst), placenta (first trimenon), placenta (mature), adrenal gland, parathyroid gland, thyroid, cerebellum, cerebrum, pituitary gland (posterior lobe), pituitary gland (anterior lobe)
In which normal tissues is the target expressed?
normal tissue analysis
140 Human Cell Lines including NCI 60
To identify tumor cell lines for functional studies/drug screening HCT-116
HCT-15HEP-G-2HT29IGR-OV1K-562LOX-IMVIMCF-7
MDA-MB-231
NCI(/L)-H226NCI-H460PC-3RPMI-8226RXF 393SF 268SK-MEL-2SK-MEL-28SK-MEL-5SK-OV-3SN 12C
SNB 19SW-620T 47 DTK 10U 251UACC-257UACC-62A 549
MDA-MB-435 (S)MOLT 4NCI-H23NCI-H322 (M)NCI-H522OVCAR-3OVCAR-4OVCAR-5OVCAR-8SF 295SF 539SNB 75
SRUO-31786-OA 498ACHNBT-549CAKI 1CCRF-CEMCOLO-205EKVX
HCC(/L)-2998HOP 62HOP 92HS-578TKM 12M-14MALME-3MKRIBT-98-GU-343-MG
LN-401LN-229BS 149
MEL HO (P4)COLO-849ECV-304CAKI-2RT-112293A 375
MBC-5/MRC-5SMBT-474(/BT-747)EAL 29
SJCRH-30WCBIM 9VM-CUB 1HELAHACATKU-19-19
GAMG p6IGR-1(/IGR 1)CRL-7930172COS-1HS-766-THUT 12HUVECIMR 90
UI-38 Mb(/U-138)
U-87 MB(/U 87 MG)WS-1HS-68MCF-10A
RT 112(/RT II2 D2I)MDA-HER-2MDANEOCAL-62DBTRGHBL-100(WBC)
Partial list
multi tumor cell line array formalin fixed
Cytospins from 33 CSC Lines
Tissue cores from 11 matched & 2 unmatched xenograftsCore diameter: 1.0mmCores per donor block: 2
Type Donors CoresGBM 8 16
Breast 1 2Thyroid 5 10Colon 7 14Lung 5 10
Melanoma 7 14Matched
xenografts: Colon 4 8Lung 3 6
Breast 1 2Melanoma 3 6
Unmatched xenografts
Breast 2 4
Total cores 92
Thyroid
Melanoma
Lung
Colon
Glioblastoma
cancer stem cell(csc) line arrayformalin fixed
2,200 Breast Cancers with 5 yr. follow-up information
pT stagepN stageNumber of nodes examinedNumber of positive nodesTumor diameterBRE gradePolymorphyTubulus formationMitoses
Tumor specific & raw survivalRadiotherapy (Y/N)
Chemotherapy (Y/N)
Molecular data: FISH: HER2, EGFR, MDM2, CCND1, MYCIHC: ERA, PR, p53, Cytokeratins, EGFR, HER2, CD117, others
breast cancer prognosis array
ESR1 Amplification* in 358/1739 (21%) of Breast Cancers
Holst, Simon et al, Nat Gen (39), 655-660, 2007
breast cancer prognosis TMA analysis
0.0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
Su
rviv
ing
0 20 40 60 80 100months surv
ER IHC positive (n=109)
p<0.0001
ESR1 amplification (n=43)
ER IHC negative(n=23)
175 Patients Treated With Tamoxifen Monotherapy
Holst, Simon et al, Nat Gen (39), 655-660, 2007
ESR1 amplification and anti ER treatment
ESR1 amplification may predict response to Tamoxifen
ABI7900 based qRT-PCR, TPD52 vs GAPDH
Normal tissues
Skin 2 Pancreas 1Lymph node 2 Stomach 2Lung 2 Kidney 2Oral cavitiy 2 Prostate 2Breast 1 Testis 3Endometrium 2 Bladder 2Ovar 2 Thyroid gland 2Vulvar 2 Brain 2Myometrium 2 Skeletal muscle 2Liver 3 Fat tissue 2
Cancers
Malignant melanoma 11 Liver cancer 50Larynx carcinoma 39 Pancreatic cancer 38Lung cancer 134 Stomach cancer 50Oral cavity cancer 56 Renal cell cancer 59Breast cancer 53 Prostate cancer 48Endometrial cancer 31 Testis cancer 59Ovarian cancer 33 Urinary bladder cancer 55Uterus cervix carcinoma 28 Thyroid gland cancer 40Vulvar cancer 39 Leiomyosarcoma 42Colon cancer 50 Liposarcoma 36Esophageal cancer 48
study: TPD52 mRNA expression analysis of 1,000 tumor samples & normal tissues
Frequency of TPD52 expression
UKE data
study: TPD52 mRNA expression analysis In 1,000 tumor samples & normal tissues
UKE data
TPD52 expression levels
study: TPD52 mRNA expression analysis in 1,000 tumor samples & normal tissues
- 5% Mutations- Mutation Unrelated To Deletion
ABI3100, 16 capillariesEppendorf pipetting robotLaser capture micro dissection (if necessary)
UKE data
tumor type data approx. # p value
PTEN not deleted 95.4% 4.6% 0.3096*
PTEN hemizygous deleted 17 11.8%FISH not
analyzable 18 0.0%
study: sequencing of all 10 PTEN exons in 100 prostate cancer samples
Breast Cancer with Herceptin Treatment & Response Information
LOCALISATION REF# ORGAN UNIQUE ID AGE DATE OF SURGERY DIAGNOSIS
GRADE T N M (TIME 0) STAGE HER2 ER (%) PR (%)METS POST SURGERY
(MONTHS)SITE OF MET. METS DIAGNOSIS BY
PREV. CHEMOTH.
SETTINGSTART
TREAT 1TREAT 1DETAILS
END TREAT 1
START TREAT 2
TREAT 2DETAILS
END TREAT 2
FOLLOW UP 1
FOLLOW UP 2
FOLLOW UP 3
SURVIVAL
sample data fields
Prospective collection of formalin fixed samples of mantle cell lymphoma from lymph node sites only with 5-10ug matching RNA per sample
Prospective collection of Frozen OCT & FFPE samples of IBD & Ulcerative Colitis, recently diagnosed, diseased + adjacent normal. Matched Serum & Whole Blood with Clinical Labs
Prospective collection of formalin fixed & OCT samples of metastatic NSCLC (adenocarcinoma & SCC) with matched nodal mets, serum & RNA
Prospective collection of formalin fixed & OCT samples of esophageal adenocarcinoma, serum & RNA
sample prospective collection projects
overview
Complexity of translational biomarker research supporting drug & diagnostic development increasingly requires knowledge-based services/partnerships that go beyond the traditional fee-for-service modelService providers must offer a range of services, histology labs, analytical platforms, top academic opinion leaders etc.
TriStar’s service platform is sustainable, scalable, flexible & cost-effectiveVery large product offering, standardized QC, top-notch scientific capabilities
contact usTriStar Technology Group LLC9700 Great Seneca Highway
Rockville, MD 20852
For more information please visit our website at www.tristargroup.us
p. 1-866-851-STAR
f. 1-509-471-1765