Patient derived phenotypic screening system utilizing Breast cancer. Offers clinical diversity met in Clinic.

1. Onco -Breast CancerPrime
TM
OncoPrime
TM
OncoPrime
TM
Use a primary cell-based platform that truly represents the clinical diversity of breast cancer - Switch to
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All Rights Reserved | Saarum Sciences Pvt. Ltd., IICT, Tarnaka, Hyderabad 500007, India. http://www.saarum.com
Figure 2. Diversity of samples available in the biobank. With steady inflow of patient
samples, we have banked a generous diversity of Breast cancer samples. The charts
represents high level categories only. Breast cancer staging as per WHO guidelines.
Triple -ve HER2+ Luminal B Luminal AMolecular
Subtypes
% 15-20% 10-15% 20% 40%
Receptor
Expression
Histologic
grade
Prognosis
Response
to therapy
Our Biobank Diversity
HER2 ER+ / PR+
High (grade III) Low (grade I)
Poor Good
Chemotherapy
Trastuzumab
Hormone Rx
Figure 1. Disease Diversity. Based on biomarker expression breast cancer can be categorized into four major subtypes. Triple negative (TNBC) tumors respond best to chemotherapy
similar to other aggressive cancers. Luminal A tumors respond best to endocrine therapy e.g. antiestrogen or aromatase inhibitor.
Patient cells characterized across several passages
“ Why use 2D cell lines when you can take advantage of
3D primary cell cultures“
Figure 3. Cells derived from patient tumours proliferate well in primary cultures
and maintain their morphology through successive generations. Representative
phase contrast images of SB-Br-523 (Infiltrating Ductal Carcinoma Grade III,
ER +ve, PR +ve, and HER2 +ve). Images were acquired with a 10X objective.
P1 P2
P3 P4
Figure 4. Human mammary epithelial cells isolated from normal breast tissue
by collagenase digestion were cultured as monolayers (Top panel) phase contrast
images shown. Bottom panel shows Mammospheres (arrows) grown in 2% agarose,
48h after passage. These 3D cell cultures are ideal for drug testing and other assays.
P1 P2
P2
100X
100X
ER/PR -ve / Her2
+ve
ER/PR +ve / Her2
+ve
ER/PR +ve / Her2
-veER+ve/PR +ve /
Her2 +ve
ER-ve /PR -ve /
Her2 -ve
Breast cancer - ER/PR/HER2 STATUS
platform is ideally suited for drug screening & lead optimization
“
Figure 5. Effect of reference compounds. Cell viability assay demonstrates effect of standard of care (SOC) compunds on three main categories of patient derived breast cancer cells
Er/Pr Negative, Her2 Positive
Log Concentration
Doxorubicin
Paclitaxel
5-FU
Cellviability
Er/Pr Positive, Her2 Negative
Doxorubicin
5-FU
Gemcitabine
Log concentration
Cellviability
Triple negative
Log Concentration
Cellviability
Doxorubicin
5-FU
27% 12% 28% 33%
Breast Cancer Stages
Our Breast Cancer Sample Collection
I A
II A
II B
III A
III B
III C

2. OncoPrime
TM
OncoPrime
TM
Onco -Breast CancerPrime
TM
A comprehensive array of phenotypic characterizations...
“
Some early evidence for Epithelial to Mesenchymal Transition (EMT) Model
“
Applications of the platform
“ Advantages of the platform
“
All Rights Reserved | Saarum Sciences Pvt. Ltd., IICT, Tarnaka, Hyderabad 500007, India. http://www.saarum.com
Figure 6. Cells of SB-Br-6170 in primary culture have a diploid amount of genomic DNA.
A high percentage of cells are arrested at the G0/G1 phase at the time of sampling.
4.84% 79.16% 4.76% 12.10%
62.85% 62.62%
48.54% 6.93% 0.06%
Figure 8. Representative images of immunofluoresence staining of SB-Br-6045 cells
in culture (ER and PR negative, positive for HER2). These cells show differential staining
for epithelial and mesenchymal markers.
Images were captured by confocal microscope with a 60X objective.
DAPI EpCAM Vimentin Merged
SB-Br-6045
Figure 7b. Human mammary epithelial cells isolated from normal breast were cultured
and stained with antibodies against EpCAM and CK 8/18. Similarly the human
mammary fibroblasts were stained with antibodies against Vimentin and alpha-SMA.
Images were captured by confocal microscope with a 40X objective.
CK 8/18EpCAM alpha-SMAVimentin
60X60X40X40XNormal Breast
• Biomarker identification and validation
• Target identification and validation
• NME / NBE / NCE screening
• Drug repurposing
• Biosimilar / Biobetter characterization
• Drug mechanism of action elucidation
• Accurately reflect the disease phenotype and diversity
• Better characterization and easy to adopt
• Cost effective and shorten time to end points
• Accelerated Preclinical Development
• Novel disease models such as EMT and MET
• 3D spheroid cultures to better mimic in vivo conditions
Morphology
Morphological evaluation at each passage
Cell surface markers – common and specific
Including sterility and Mycoplasma testing
Mutation and Epigenetic Profiling - Breast cancer
BRCA1/2, EGFR, B-RAF, K-Ras, HER-2/Neu, TP53,
C-Met, C-myc, PI-3K, MEK, FLT-3, PDGFR, TERT1/2
Functional assays - Reference drug data
Cell migration and invasiveness
Colony formation and sphere formation
Standard of care (SOC) data for drug screening
Regulated model of Epithelial Mesenchymal Transition
Target discovery or validation,
ID & screening of new drugs / combinations
ID novel EMT / Metastasis biomarkers
Therapeutics targeting EMT will target the crucial step of
- metastasis and the formation of secondary sites of cancer.
Day 0 Day 7 Day 11
Day 12 Day 14 Day 15
35 mm 35 mm 35 mm
48 well 48 well 24 well
Figure 7a. Top Panel: Epithelial to mesenchymal transition after transfection of
primary normal breast epithelial cells with our provisional patented constructs.
Bottom Panel: At Passage 1, induced cells show positive staining (+++) for
mesenchymal markers and minimal staining (+) for epithelial marker which
is reversed by Passage 2. DAPI - blue, CK 8/18 & EpCAM - green, SMA - Red
DAPI, EpCAM & Vimentin
DAPI, CK 8/18 & SMA