Академический Документы
Профессиональный Документы
Культура Документы
Faculty
Michael F. Press, MD, PhD
Harold E. Lee Chair in Cancer Research Professor of Pathology Department of Pathology University of Southern California, Norris Comprehensive Cancer Center Los Angeles, California
Disclosure
Michael F. Press, MD, PhD, has disclosed that he has received consulting fees from GlaxoSmithKline, Halozyme, and Roche and funds for contracted research from Genentech and Ventana Medical Systems.
17
17
Southern
(DNA)
4.4 kb p185 -
Northern
(mRNA)
Western
(protein)
Frozen IHC
27%
Slamon D, et al. Science. 1989;244:707-712.
63%
% Women
10%
Amplification Level :
> 10 5 - 10 2-5 1
IHC
H&E
Southern blot
FISH
Chromosome 17 centromere
Interpretation Signal enumeration Ratio of HER2:Chr 17 signals
2+
3+
Evaluation of HER2 Testing in Paraffin-Embedded Tissues Correlation of HER2/neu Gene Amplification With Overexpression in Frozen and FFPE Tissues
Amplification Level : 12.5 kb -
> 10 5 - 10 2-5 1
Southern
(DNA) 4.4 kb p185 -
Northern
(mRNA)
Western
(protein)
Frozen Tissue
Slamon D, et al. Science. 1989;244: 707-712.
27%
63%
Press MF, et al. Cancer Res. 1993;53:4960-4970. Press MF, et al. Cancer Res. 1994;54:2771-2777 Press MF, et al. J Clin Oncol. 2002;20:3095-3105.
Selection of Assay Method for Clinical Trials Samples Comparison of FISH and IHC Assay Results in Cancers With Known HER2/neu Gene Status
Assay
FISH (Vysis) FISH (Ventana) IHC-R60 IHC-10H8
#Amplified or #Over/#Low
41/76 44/73 39/78 38/79
Sensitivity
95% 95% 91% 88%
Specificity
99% 96% 100% 100%
Accuracy
97% 96% 97% 96%
IHC-DAKO IHC-Ventana
30/87 31/86
70% 72%
100% 100%
89% 90%
1+
Positive for HER2 protein expression IHC 3+ (defined as uniform intense membrane staining of > 30% of invasive tumor cells)
Equivocal HER2 gene amplification (patients with HER2/CEP17 ratio 2.0 were eligible for the adjuvant trastuzumab trails)
CTA-IHC 1+ 2+ 28 67
3+
21
7
3%
2
7%
21
24%
176
89%
Amplification rate
3.7%
Overall concordance between FISH and IHC results was 82% (95% CI: 78%-85%; P < .0004)
Dybdal N, et al. Breast Cancer Res Treat. 2005;93:3-11.
Local IHC Compared to Central FISH (79% Concordance) Central Screening of HER2 Status for Entry to Adjuvant Trastuzumab Trial (BCIRG006) Local IHC 0 1+ 2+ 3+ 538 20 4% 230 15 6% 4.3%
Press MF, et al. Clin Cancer Res. 2005;11:6598-6607.
165 33 17%
90 316 78%
73%
Central FISH
35/384 = 9.1% +
23%
Amplification rate
N = 1407
Overall concordance between FISH and IHC was 79% (95% CI: 77%-81%)
Prevalence of HER2 Gene Amplification (%) in Each IHC Immunostaining Category (0, 1+, 2+ and 3+) by Study*
HER2 Gene Amplification Rate according to IHC Score 0 1+ 2+ 3+ Number IHC Method Reference
0%
1.8% 3.5% 0% 0% 3.8% 0% 0.7% 4.2% 1.1% 0%
0%
17%
35.9%
89%
100% 99% 91.4% 100% 100% 89.8% 94.1% 49% 89.7% 90%
100
750 2857 189 170 198 119 426 102 2279 360
DAKO HercepTest
DAKO Ab, Unspecified DAKO HercepTest DAKO A0485 Ab Homebrew Ab DAKO HercepTest DAKO HercepTest DAKO HercepTest DAKO HercepTest DAKO HercepTest DAKO HercepTest
3.1%
26.5% 20%
0%
2.8% 3% 6.9% 2.4% 3.6% 12.5% 0% 2.7% 8.7% 6.1% 6.7% 0% 21.4% 7%
15%
17% 24% 31.8% 72% 16.7% 7% 12.2% 36.1% 36.1%
79%
91.6% 89% 90% 100% 78.1% 52.4% 91.6% 90.1% 90.1%
600
2913 529 114 215 2249 108 289
DAKO HercepTest
DAKO A0485 Ab Clinical Trials Assay DAKO HercepTest DAKO HercepTest DAKO HercepTest DAKO HercepTest DAKO HercepTest
Prevalence of HER2 Gene Amplification (%) in Each IHC Immunostaining Category (0, 1+, 2+ and 3+) by Study (2)*
HER2 Gene Amplification Rate according to IHC Score 0 1+ 2+ 3+ Number IHC Method Reference
0%
1.60% 12.5% 3.3% 0% 2.6%
8.30%
23%
56.3%
661
DAKO HercepTest
34.29%
68.6%
57.9% 3.30% 4.80% 15.20% 28.10%
86.00%
96.3%
95.2% 84.1% 93.8%
697
171
100 200 950
0%
12.8%
3.17%
21.51%
43.8%
90.98%
97.8% 100.0% 88.4% 97.3%
89.65%
681
291 216 543 125
4635
Dako Herceptest
0% 3.4% 0%
Martin et al, Patholog Res Int., 2012 Lee et al, Arch Med Res., 2012 Kiyose et al, Pathol Int., 2012
0.86%
FDA approved a HER2-toCEP17 ratio 2.0 (or an average HER2 copy number of > 4.0) as HER2 amplified
Negative for HER2 gene amplification (FISH ratio > 1.8 or HER2 gene copy < 4.0)
Positive for HER2 gene amplification (FISH ratio > 2.2 or HER2 gene copy > 6.0)
Equivocal for HER2 gene amplification (FISH ratio 1.8-2.2 or HER2 gene copy 4.0-6.0*)
Count additional cells for FISH or retest, or test with HER2 IHC
Equivocal HER2 gene amplification result (patients with HER2/ CEP17 ratio 2.0were eligible for the adjuvant trastuzumab trails
No targeted therapy
ASCO-CAP guidelines create an equivocal zone (1.802.20) without providing any supportive data Considerable data and the FDA consider this zone to be the area around the cut-off of amplification where additional cells should be scored
HER2 FISH Ratios in 2502 Consecutive Cases Screened for Entry to Clinical Trials: Equivocal Cases Only a Minute Assessment Problem
10.00 9.00 8.00 HER2 FISH Ratios 7.00 6.00 5.00 4.00 3.00
2.20
1.80
Cases
Correlation of PFS With Response to Lapatinib Based on HER2 FISH Status in HVLab vs Academic Lab
Blinded FISH analysis by large, commercial, high-volume laboratory
Number 255 86
HR 0.47 0.54
Positive Negative
271 47
0.46 0.94
0.33-0.65 0.39-2.28
HER2 FISH Ratios in 2502 Consecutive Cases Screened for Entry to Clinical Trials
Plot of FISH Ratios 10.00 9.00 8.00
1.80
HER2 Gene Amplification by FISH Compared With HER2 Protein by IHC IHC-HercepTest
0 1+ 2+ 3+
237 13 5%
99 21 21 18% 9%
36 39 52%
8 258 97%
53%
47%
Amplification rate
N = 711
Overall concordance between FISH and IHC results was 89% (95% CI: 87%-91%).
P < .0001
Press MF, et al. Clin Cancer Res. 2008;14:7861-7870.
HER2 IHC 3+
Improved Outcome Is Associated With Entire Spectrum of HER2 Amplification Ratios (FISH)
FISH Ratio < 2.0 2.00-4.99 5.00-6.99 7.0-10.0 10.0 Number 390 59 89 106 92 HR 1.09 0.44 0.45 0.58 0.39 95% CI 0.86-1.37 0.24-0.83 0.25-0.81 0.33-1.02 0.22-0.68 P Value .486 .010 .007 .059 .001
Analysis of all cases in the ALab for patients in either clinical trial.
Low Level HER2 Gene Amplication Appears to Respond to Trastuzumab Treatment: FISH Ratio 2.0-2.2
100 90 80 DFS (%) 70 60 50 40 0 1 2 3 4 5 Time From Registration (Yrs) 6 FDA only ACT Ref ACT+H HR: 0.60; 95% CI: 0.12-3.13; P = .55 ASCO/CAP ACT Ref ACT+H HR: 0.59; 95% CI: 0.48-0.73; P < .001
ClinicalThought an interactive forum where you can discuss the latest trends in breast cancer with experts
clinicaloptions.com/oncology