Gene expression profiling in breast carcinoma
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Transcript of Gene expression profiling in breast carcinoma
GENE EXPRESSION PROFILING IN BREAST CARCINOMADr Partha Ghosh2nd year PGT, General SurgeryN.R.S Medical college
What is gene expression profiling?Cancer is a disease characterized by uncontrolled
cell growth and proliferation. For cancer to develop, genes regulating cell growth and differentiation must be altered; these mutations are then maintained through subsequent cell divisions and are thus present in all cancerous cells. Gene expression profiling is a technique used in molecular biology to query the expression of thousands of genes simultaneously. In the context of cancer, gene expression profiling has been used to more accurately classify tumours. The information derived from gene expression profiling often has an impact on predicting the patient’s clinical outcome.
Tumor biology
IHC mRNA profiling Histology TNM Staging
Breast Cancer SubtypesBreast cancer is classified into clinical
subtypes based upon receptor expressionThese subtypes dictate possible therapeutic
options and vary in their prognosis◦ Luminal: derived from the luminal cells
TYPE A:ER+, PR+,HER-2- TYPE B:ER+,PR+,HER-2+ Can use hormonal therapy Less aggressive
◦ Basal: derived from myoepithelial cells ER-, PR-,HER-2-,ck 5/6+ or HER -1+ No specific target for therapies More aggressive
◦ HER2-enriched More aggressive
Luminal A
Luminal B
Claudin-Low
HER2-enriched
Basal
Luminal and Basal Characteristics
BasalLow ERLow HER2High CK5/6c-KIT higherHigh EGFRHigh p53
mutationHigh p53 proteinHigh cyclin EVery high
vimentin
LuminalHigh ERHigher HER2Low CK5/6Low c-KITLow EGFRLow p53
mutationLow p53 proteinLow cyclin ELow vimentin
Basement membraneMyoepithelial Cells BasalLuminal Cells Luminal
What is HER 2/neuOther names
◦Receptor tyrosine-protein kinase erbB-2
◦ CD340 ◦ proto-oncogene Neu◦ ERBB2 (human)Located in chromosome 17 (17q12)Rodent glioblastoma cell line a neural
tumour
Cont….Plasma membrane bound receptor
tyrosine kinasePositive in 15-30% cases of breast
carcinomaAssociated with increase disease
recurrence and poor prognosisTested by-
◦Immunohistochemistry ◦FISH◦Serum HER2 by ELISA- Trastuzumab
response
Cont…Drugs targeting HER2
Trastuzumab- p27 halts cell proliferatonPertuzumab- prevent dimerisation of HER2 and HER3NeuVax- Direct killer T cells to destroy HER2 +ve cancer cells
Tamoxifen resistant breast caThe expression of HER2 is
regulated by signaling through estrogen receptors.
Normally tamoxifen down regulate HER2
Ratio of coactivator AIB-3 exceed that of corepressor PAX2 HER2 expression in presence of tamoxifen
Algorithm for breast cancer subtypes
All cases
ER - PR -ER+ or PR +
HER2 - HER2 + HER2 +
HER2 -
EGFR - CK5/6 – EGFR + or CK5/6 +
Unclassified Basal Like Luminal B Luminal AHER2+/ER-
Luminal ALuminal B
HER2+Basal-like
Intrinsic Breast CancerSubtypes described by
Perou et al.
Express ↑ amountsOf luminal cyto-Keratins & geneticMarkers of luminalEpithelial cells ofNormal tissue
Express ↑ levels of EGFR, c-kit, & growth factors like hepatocyte growth factor and IGF
Mammaprint: Development of the 70-Gene Signature DNA microarray analysis of 78 breast primary tumors
(untreated) Pts were <55 years of age with T1-2/N0 disease Pts selected based on outcome: Distant metastases
within 5 years
Statistical analysis, “supervised classification,” identified 231 genes correlated with disease outcome Top 70 genes selected
Genes that regulate cell cycle, invasion, metastasis, & angiogenesis
Patients categorized as “good prognosis” or “poor prognosis.”
Found to be a better predictor of distant metastases within 5 years than all clinical variables in this study
Odds ratio (distant metastases): poor to good prognosis groups = 15
Does 70-gene Signature have Independent Prognostic Value?
Gene signature adds independent prognostic information to that provided by various risk classifications
The signature remained a statistically significant prognostic factor for time to distant metastases & OS even after adjustment for various risk classifications (HR 2.15 & 2.15, respectively)
The 21-gene Recurrence Score: Oncotype DX
Oncotype Dx: The 21-Gene Assay Designed to quantify the risk of distant recurrence in
patients with LN(-), ER(+) tumors receiving tamoxifen RT-PCR was used to quantify gene expression from
fixed, parafin-embedded tumor tissue 250 candidate genes selected based on published
literature, genomic databases, & experiments based on DNA arrays on fresh-frozen tissue
Analyzed data from 3 independent studies (447 patients) including tamoxifen-only arm of NSABP trial B-20 to test relation b/w 250 genes and recurrence of breast cancer
From these studies, 16 genes (+5 reference genes) were selected that correlated with proliferation and endocrine response
Levels of Gene Expression Determine Recurrence Score
21-gene assay = 16 outcome-related genes + 5 reference genes
Higher expression levels of“favorable” genes = ↓ RS
Higher expression levels of“unfavorable” genes = ↑ RS
A risk score is calculated from 0 -100Cutoff points chosen based onResults of NSABP trial B-20
Prospective Validation of Oncotype DX:TAILORX Trial
Low RS:HormonalTherapy
High RS:Chemo +HormonalTherapy
Hormonal Therapy Chemo + Hormonal
11,248 ER+/LN- patients
TN VS BASAL SubtypesTriple negative
◦ ER,PR,HER2 receptor negative
Basal Subtypes◦ ER,PR,HER2
receptor negative◦ Over expression
of breast basal cell epithelium gene
◦ EGFR+,CK5/6+
Basal like subtypes of breast cancerNo proven therapeutic target
ER,PR negative-cannot use tamoxifen or anti estrogen
HER2 negative-cannot use Herceptin
Targeted therapy ER + ER-
Hormonal Herceptin EGFR BRCA 1 C-kit
Therapy Geftinib, erlotinib, DNA damage Imatinib
lapatinib PARP inh
Luminal A
Luminal B HER 2 + Basal
Like
Potential treatment options in TNBC
Taxane and Anthracycline Based TherapyTypical regimens:
AC-T: doxorubicin plus cyclophosphamide every 2 weeks for four cycles followed by docetaxel every 2 weeks for 4 cycles
TAC: docetaxel, doxorubicin, and cyclophosphamide every 3 weeks for 6 cycles
Dense dosing is better
Platinum AgentsPlatinum agents can bind to DNA
and cause cross-linking to occur cell death
Cisplatin, carboplatin and oxaplatin are approved for some types of cancers and are being studied as treatments for TNBC
PARP InhibitorsPARP: poly ADP ribose
polymerase◦Involved in DNA repair by detecting
single-strand breaks◦Can be activated in cells with
damaged DNASeveral types of cancer are more
dependent on PARP, so it can be a good therapeutic target
PARP inhibitors prevent breaks from being repaired, causing cell death
Anti-EGFREGFR is overexpressed in 45-70% of TNBC Cetuximab is an anti-EGFR antibody used to
treat metastatic cancer◦Breast cancer patients with metastatic disease
respond twice as well when Cetuximab is addedOther treatments include tyrosine kinase
inhibitors (erlotinib, gefitinib)◦Gefitinib is the only one currently approved for
breast cancer, but the others are in clinical trialsInhibits an important signaling pathway and
provides a specific target!
Angiogenesis in CancerAngiogenesis: formation of new
blood vessels. ◦Tumors need blood vessels to grow and
spread.Angiogenesis inhibitors prevent the
formation of new blood vessels, thereby stopping or slowing the growth or spread of tumors.
Anti-AngiogenesisBevacizumab (Avastin)
◦ Monoclonal antibody to VEGF◦ Improves survival in breast cancer patients with
combined with Taxol◦ Approved for metastatic breast cancer but benefit
isn’t subtype specific – this has since been revoked because it slowed progression but didn’t extend length or quality of life and had many adverse effects
Metronomic chemotherapy: repeated, low, less than toxic doses can destroy endothelial cells and prevent angiogenesis, slowing tumor growth – works in clinical trials
Androgen ReceptorNuclear receptor activated by binding
testosterone or dihydrotestosterone◦ Closely related to PR
Expressed in 75% of breast cancer and 10-20% of TNBC◦ TNBC that express AR are molecularly similar to
prostate cancer and could potentially be treated similarly.
Bicalutamine: anti-androgen used to treat prostate cancer
17-DMAG: semi-synthetic antibiotic derivative, has shown promise in clinical trials
Enzalutamide: androgen agonist used to treat prostate cancer; is in Phase II for TNBC
RTK InhibitorsSuninitib (Sutent)
◦Multiple-target RTK inhibitor All PDGFRs and VEGFRs KIT (CD17) which drives the majority of all GI
stromal tumors & several others
Imatinib (Gleevec)◦Prevents phosphorolation of BCR-Abl,
inhibiting signaling pathways necessary for cancer cell growth BCR-Abl: Exists only in cancer cells! Worked in vitro; no effect on metastatic
breast cancer patients in Phase II
Src Tyrosine KinaseSrc is overexpressed in breast
cancerDasatinib: multiple tyrosine kinase
inhibitor approved for CML◦Possible efficacy in breast cancer -
small effect seen in Phase II◦In vitro: basal breast cancer cells were
more sensitive!Several others in trials also seem to
have promising preclinical activity
mTORCell cycle regulator and a downstream
effector in the PI3K/PTEN/AKT pathwayPTEN is often mutated in TNBC, leading
to increased AKT and mTOR activationEverolimus and temsirolimus block mTOR
function and inhibit proliferation◦Everolimus is approved for some types of
cancers - currently in clinical trials for TNBC in combination with chemotherapy
◦Temsirolimus is approved for renal cell carcinoma and completed a Phase II trial with promising results
ConclusionsGene signatures augment current
clinicopathological variables in assessing risk of recurrence
Gene expression profiles may be both prognostic and predictive for patients with early breast cancer
NCCN guidelines suggest that Oncotype DX is an option for risk evaluation in 0.6-1 cm tumors with unfavorable characteristics or in >1 cm LN-, ER+/HER2 negative tumors
NCCN guidelines include Oncotype DX® testing in the treatment-decision pathway for node-negative and micrometastatic disease
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• Tumor 0.6-1.0 cm, moderately or poorly differentiated, intermediate or high grade, or vascular invasion
• Tumor > 1 cm with favorable or unfavorable pathologic features
Consider Oncotyp
e DX
Hormone receptor-positive, HER2-negative disease
pT1, pT2, or pT3 and pN1mi
No test
RS < 18
RS 18-30
RS ≥ 31
Adjuvant endocrine therapy
± adjuvant chemotherapy
Adjuvant endocrine therapy
endocrine therapy ± adjuvant chemotherapy
Adjuvant endocrine therapy
+ adjuvant chemotherapy
THANK YOU