Table 6.
Studies evaluating relationship between topoisomerase IIa and response to anthracycline vs non-anthracycline or differing doses of anthracyclines.
| Study | Publication | Regimens | Setting | N | Outcomes | Topoisomerase 2a test/definitions |
|---|---|---|---|---|---|---|
| Belgian Di Leo 2001 | 77 | CMF vs EC vs High dose EC | ADJ | 481 |
Most samples had 1–25% of cells positive for TopoIIα. 161 tumors TopoIIα positive; only 25 were HER2 positive by IHC TopoIIα expressing trended toward improvement in EFS with an anthracycline-based adjuvant therapy. 50 TopoIIα + High-EC vs CMF: HR = 0.66, p = 0.25; 50 TopoIIα- High-EC vs CMF: HR = 1.26, p = 0.51; p interaction 0.13 |
IHC only KiS1 Ab >10% is positive HER2 FISH >2 = positive |
| Belgian Di Leo 2002 | 78 | CMF vs EC vs High dose EC | ADJ | 61 |
23/61 of HER2+ were TOP2A+ (did not look for TOP2A in HER2 negative) Trend toward EFS benefit with EC or High dose EC only in HER2+ tumors Co-amplified had better EFS with EC or High dose EC vs CMF; TOP2A negative did not appear to benefit from anthracyclines |
FISH HER2+ ratio >2.0 TOP2A+ ratio >1.5 TOP2A ratio <0.8 del |
| Belgian Case-Control Cardoso 2004 | 132 | Anthracycline vs taxane | MBC | 58 |
TOP2A amplification in eight tumors. All were HER2+ TOP2A+ treated with anthracycline: 21% CR; 6% progressive disease; TOP2A+ treated with taxane: 14% CR and 14% progressive disease |
FISH HER2 ratio >2.0 TOP2A ratio >1.5 positive IHC >10% |
| DBCG 89D* Knoop 2005 | 81 | CEF vs CMF | ADJ | 805 |
246 HER2 positive by IHC or FISH; 79 of these (32%) TOP2A co-amplified. 14/527 HER2 negative (IHC), TOP2A amplified Anthracycline benefit similar in HER2+ and HER2−. TOP2A amplification had improved RFS (HR = 0.43) and OS (HR = 0.57) with CEF. TOP2A del may also associate with outcome. |
FISH HER2 and TOP2A ratio >2 positive TOP2A ratio <0.8 del |
| DBCG 89D* Nielsen 2008 | 146 | CEF vs CMF | ADJ | 767 |
Updated analysis of TOP2A aberrations with IVD-labeling; TOP2A status/N: Amplified: N = 92 (12%); Deletion 87 (11%) Normal: N = 589 TOP2A normal have longer RFS than TOP2A amp or del RFS CEF vs CMF: TOP2A amplified: HR = 0.39, p = 0.002; TOP2A deleted: HR = 0.61, p = 0.083; TOP2 negative: HR = 0.94, p = 0.60 OS CEF vs CMF: TOP2A amplified: HR = 0.48, p = 0.01; TOP2A deleted: HR = 0.68, p = 0.155; TOP2 negative CEF vs CMF: HR = 0.90, p = 0.42 |
FISH HER2 and TOP2A ratio >2 positive TOP2A ratio <0.8 del IVD-labeling of TOP2A FISH pharmDx™ Kit |
| DBCG 89D* Ejlertsen 2010 | J Clin Oncol 2010;28:984-90 | CEF vs CMF | ADJ | 623 | Looked at TIMP1, HER2 and TOP2A and association with anthracycline benefit. Combining TOP2A abnormal with TIMP1 negative expression (2T responsive profile) predicted better iDFS and OS with CEF vs CMF. Stronger predictor than HER2+ ± TIMP1−. underpowered |
FISH HER2 and TOP2A ratio >2 positive TOP2A ratio <0.8 del |
| Scandinavian Breast Group 9401 Tanner 2006 | 174 | FEC × 9 (Tailored/dose escalated) vs FEC × 4 followed by ASCT (less anthracycline) | ADJ | 391 of 525 |
HER2 by CISH, TOP2A checked in HER2+ (N = 128) only Significantly shorter RFS and OS in HER2+; 37.5% (48/128) TOP2A coamplified. Patients with TOP2A-amplified tumors had a statistically significantly better breast cancer RFS when treated with tailored and dose escalated FEC (high dose anthracycline) compared with standard FEC-ASCT (HR = 0.45; P = 0.049); no such difference between FEC9 and ASCT in patients with no TOP2A amplification (HR = 0.95; P = 0.88). Multivariate cox regression analysis showed among TOP2A amplified, FECx9 treatment was the only significant predictor for a better outcome (adjusted HR = 0.30; P = 0.020). For tumors without TOP2A amplification, treatment did not predict outcome in multivariate analysis. |
CISH Ratio > 2 or >6 copies in >30% of nuclei or presence of an easily identifiable gene copy cluster, if gene copies could not be counted |
| TAX303 Durbecq 2004 | 164 | A vs T | MBC | 108 |
MBC: 43 patients Topo2 protein positive, higher response to anthracyclines if Topo2+. No diff in TTP and OS Both 2004 and 2007 analyses show similar results by expression |
IHC: TopoII α-positive: >10% of immunostained cells |
| TAX303 Di Leo 2007 | 165 | A vs T | MBC | 91 |
36/91 TOP2A+; The probability of response to doxorubicin is higher in topoII α-positive than in topoII α-negative tumors [Odds ratio 4.51 (95% CI 1.01–20.10), P = 0.05]. This correlation is not found for docetaxel [OR 0.75 (95% CI 0.21–2.75), P = 0.66]. Response to anthracycline is highest (5x better) for p53 wild type/Topo2a+ (N = 16) vs other pts |
See above |
| MA-5 O’Malley 2009 | 149 | CEF vs CMF | ADJ | 438 (62%) |
33/116 HER2+ TOP2A+ (28%); 20/314 HER2 neg, TOP2A amp 12% (54/438); TOP2A deletion 6% (26/438) TOP2A amplification trended toward a relationship with better RFS and OS with CEF than CMF but not statistically signif. 5-year RFS CEF vs CMF TOP2A+ (N = 54): HR = 0.51, p = 0.20; TOP2A del (N = 26): HR = 0.16, p = 0.02; TOP2A normal (N = 358): HR = 0.90, p = 0.49; p interaction 0.22 5-year OS CEF vs CMF TOP2A+: HR = 0.47, p = 0.20; TOP2A del: HR = 0.18, p = 0.07; TOP2A normal: HR = 1.09, p = 0.62; P interaction 0.07 After adjustment, HR interaction between treatment and altered vs normal TOP2A 0.53 for RFS (P = 0.09) and 0.38 for OS (P = 0.02). |
FISH HER2 and TOP2A ratio >2 positive TOP2A ratio <0.8 del |
| BR9601 Bartlett 2008 | 84 | CMF vs E-CMF | ADJ | 303 |
No association between TOP2A amplification or alteration in RFS and OS; No association between TOP2A amp/alteration and outcome with anthracycline. Definition of TOP2A amplification different from HER2. 63 HER2 amplified; worse RFS and OS with epirubicin 26/303 TOP2A amplified (9%), 17/26 coamplified for HER2; nine TOP2A+, HER2 Nl (may be due to TOP2A amplification definition) TOP2A deletion 50/303 (17%) – did not evaluate outcome or interaction based on this alone |
FISH: HER2 ratio >2.0; TOP2A ratio >1.5 positive TOP2A ratio <0.8 del IHC: >median |
| BR9601/NEAT Bartlett 2010 | 85 | CMF vs E-CMF | ADJ | 1762 |
367 HER2 amplified (21%); 169/1762 TOP2A amplified (10%); 191/1762 TOP2A deleted (11%). Unknown how many are co-amplified. -HER2 amp or TOP2A del worse RFS/OS. TOP2A amp and CEP17 dup no relation to RFS/OS. -no significant interaction with anthracycline benefit for HER2 or TOP2A (del or amplification or combined). -CEP17 duplication predicted RFS and OS benefit from anthracycline (significant interaction) HER2 amp and TOP2A alterations (amp and del) significantly associated with CEP17 dup (P < 0.0001) |
HER2 ratio >2.0 (different from above) TOP2A ratio >1.5 TOP2A ratio <0.8 del CEP17 dup = >1.86 signals/cell |
| CALGB 8541 Harris 2009 | 143 |
CAF Different doses |
ADJ | 624 of 1572 |
HER2+ 117/624 = 19% TOP2A amp 41 (7%), del 69 (11%); 39/41 HER2/TOP2 co-amplified TOP2A amp does not predict benefit from CAF in HER2+. 41 patients TOP2A+ spread over three arms; All pts received anthracycline. No info regarding the distribution of patients across dose strata (very small numbers in each of three dose strata, underpowered). |
FISH HER2 & TOP2A Ratio >2 = positive Ratio <0.67 = del |
| H0648 Press 2011 | 136 | Anthracycline or taxane ± trastuzumab | MBC | 339 of 469 |
279 confirmed to be HER2 amplified; 99/279 coamplified TOP2A (35% of HER2+); 45/279 (16%) del TOP2A in HER2+ No TOP2A amplification in HER2 negative. 2/60 HER2 neg, TOP2A deleted 2/60 HER2 negative. TOP2A/HER2 coamplified: longer PFS/OS compared with normal TOP2A if treated with anthracycline. In non-trastuzumab treated patients: OS better with anthracycline if TOP2A amplified (38.5 mos) vs non-amp (18.2 mos, p = 0.004) and OS same with paclitaxel for TOP2A amp (18.4 mos) vs non-amp (20.6 mos) (p = NS) |
FISH HER2 and TOP2A Ratio >2 = positive; Ratio <0.82 = del Used methods and probes validated from physical mapping of 17q12-q21 amplicon. |
| BCIRG006 Press 2011 | 136 | AC-T vs AC-TH or TCarboH | ADJ | 2990 |
1057/2990 (35%) HER2/TOP2A coamplified; 145/2990 HER2+/TOP2 deleted If TOP2A amplified, AC-T DFS/OS improved similar to AC-TH and TCH. If TOP2A non-amplified, AC-T does worse, AC-TH = TCarboH |
FISH HER2 &TOP2A Ratio >2 = positive; Ratio <0.82 = del |
| Meta-analysis (Belgian, MA-5, DBCG89b, NEAT, BR9601) Di Leo 2011 | 87 | CMF vs anthracycline | ADJ | 3102 |
3102 tested for TOP2A, 3452 for HER2. 316/3102 (10%) deleted for TOP2A, 275 (9%) TOP2A amplified. Concordance of 123 samples tested at external lab (Tampere) compared to results from university labs: 69% Did not provide % of pts with HER2/TOP2A co-amplification or TOP2A amp in HER2 negative EFS anthracycline vs CMF: TOP2A normal: 0.88 (0.78, 1.00); TOP2A del: 0.63 (0.46,0.87); TOP2A amp: 0.62 (0.43,0.90); p interaction = 0.0513 OS anthracycline vs CMF: TOP2A normal: 0.89 (0.78, 1.03); TOP2A del: 0.68 (0.49,0.95); TOP2A amp: 0.67 (0.46,0.98) p interaction = 0.1608 when combined deleted/amplified it is statistically significant |
FISH Ratio >2 = positive Ratio <0.8 = del |
| Meta-analysis (see above) | 134 | CMF vs anthracycline | ADJ | 3846 |
HER2: Total 3436, 832+ (24%); TOP2A: Total 3098, 273+ (9%), 314 Del (10%) CEP17: Total 3225, 971 duplicated (30%) TOP2A del and TOP2A del/amp poorer RFS and OS; Noted heterogeneity for the prognostic impact of some markers across studies Interaction for treatment with HER2 not significant for RFS/OS in adjusted analyses. Stratified by trial, adjusted for confounders, Cox regression model shows interaction for CEP17 dup and anthracycline significant for RFS (0.01), borderline for OS (p = 0.06). TOP2A interaction significant for both RFS (p = 0.03) and OS (p = 0.03) |
FISH: Ratio >2 = TOP2A and HER2 positive Ratio <0.8 = TOP2A del Definition CEP17 dup: >1.86 copies per cell for DBCG, BR9601, NEAT, Belgian; >2.25 copies/cell for MA-5 |
| TRYPHAENA Schneeweiss 2014 | Br Ca Res 16:R73 | FEC/THPer vs FECHPer-THPer vs TCarboHPer | NAC | 192 | TOP2A amplification (62/192, 32%) not associated with different pCR for FECHPer/THPer; TCarboHPer or FEC-THPer. | FISH: Ratio >2.0 = positive |
A doxorubicin, Ab antibody, ADJ adjuvant, amp amplification, BCIRG Breast Cancer International Research Group, CALGB Cancer and Leukemia Group B, C cyclophosphamide, Carbo carboplatin, CEP17 chromosome enumeration probe 17, CISH chromogenic in situ hybridization, CR complete response, DBCG Danish Breast Cancer Cooperative Group, del deleted, DFS disease-free survival, dup duplicated, E epirubicin, ER estrogen receptor, EFS event-free survival, F fluorouracil, FISH fluorescent in situ hybridization, FFPE fresh frozen paraffin embedded, HD high dose, HR hazard ratio, IHC immunohistochemistry, M methotrexate, MBC metastatic breast cancer, NAC neoadjuvant chemotherapy, NR not reported, PFS progression-free survival, OS overall survival, pCR pathologic complete response, Per pertuzumab, p int p value interaction test, RFS relapse-free survival, T docetaxel, TTP time to progression.