Prognostic Value of the TP53 Mutation Location in Metastatic Breast Cancer as Detected by Next-Generation Sequencing

Figures & data

Figure 1 Flowchart of patient inclusion.

Table 1 Baseline Clinical Characteristics of TP53 Wild-Type and -Mutated Metastatic Breast Cancer Patients (n=187)

Characteristics	TP53 Status		p value
	Wild-Type (n=108)	Mutated (n=79)
Age at diagnosis (years)
Median age(range)	46(26–80)	48(27–69)
≤50	64(59.3%)	49(62.0%)	0.702
>50	44(40.7%)	30(38.0%)
Family history
Breast/ovarian cancer	7(6.5%)	3(3.8%)	0.688
Other cancers	17(15.7%)	14(17.7%)
No	84(77.8%)	62(78.5%)
Stage at diagnosis
I~II	52(48.1%)	29(36.7%)	0.136
III	22(20.4%)	22(27.9%)
IV	18(16.7%)	20(25.3%)
Unknow	16(14.8%)	8(10.1%)
Grade
I–II	53(49.1%)	41(51.9%)	0.563
III	13(12.0%)	13(16.5%)
Unknow	42(38.9%)	25(31.6%)
Ki67
1~20%	35(32.4%)	35(44.3%)	0.245
>20%	55(50.9%)	38(48.1%)
Unknow	18(16.7%)	6(7.6%)
HR status
Positive	85(78.7%)	53(67.1%)	0.074
Negative	23(21.3%)	26(32.9%)
HER2 status
Positive	15(13.9%)	21(26.6%)	0.030
Negative	93(86.1%)	58(73.4%)
Lymph node status
Positive	65(60.2%)	44(55.7%)	0.554
Negative	42(38.9%)	34(43.0%)
Unknow	1(0.9%)	1(1.3%)
Disease involvement
Visceral	64(59.3%)	46(58.2%)	0.848
Non-visceral	42(38.9%)	32(40.5%)
Unknow	2(1.9%)	1(1.3%)
Adjuvant endocrine therapy
Tamoxifen/toremifene	41(37.9%)	23(29.1%)	0.238
Aromatase inhibitor	14(13.0%)	10(12.6%)
Tamoxifen+ aromatase inhibitor	3(2.8%)	3(3.8%)
Others	1(0.9%)	1(1.3%)
No	49(45.4%)	42(53.2%)
Adjuvant chemotherapy
Paclitaxel	9(8.4%)	12(15.2%)	0.134
Anthracycline	23(21.3%)	7(8.9%)
Paclitaxel+ anthracycline	36(33.3%)	25(31.6%)
Others	4(3.7%)	4(5.1%)
No	36(33.3%)	31(39.2%)
Adjuvant targeted therapy
Yes	2(1.8%)	4(5.1%)	0.223
No	99(91.7%)	73(92.4%)
Unknow	7(6.5%)	2(2.5%)

Abbreviations: HER2, human epidermal growth factor receptor 2; ER, estrogen receptor; PR, progesterone receptor; DFS, disease free survival.

Figure 2 The mutational spectra of TP53 in TP53-mutated patients. (

) Missense (

) Truncating (

) Inframe.

Notes: According to http://www.cbioportal.org, truncating mutations included nonsense mutations, frameshift mutations and splicing mutations.

Figure 3 Survival analyses by Kaplan–Meier according to TP53 status in MBC patients. (A and B) TP53 wild-type patients had a significantly better clinical outcome than TP53-mutated patients. (C and D) there were no significant differences between TP53 wild-type and -mutated patients in the HER2-positive cohort. (E and F) TP53 wild-type patients had a significantly longer median DFS and OS than TP53-mutated patients in the HR+/HER2– cohort. (G and H) TP53 wild-type patients had a significantly longer median DFS than TP53-mutated patients in the TNBC cohort.

Figure 4 Survival analyses by Kaplan–Meier according to TP53 mutation sites in MBC patients. (A) Patients with a mutation in the non-DNA binding domain had a significantly shorter median DFS than TP53 wild-type patients and those with mutations in the DNA-binding domain. (B) Patients with a mutation in the non-DNA binding domain had shorter median OS than TP53 wild-type patients and those with mutations in the DNA-binding domain. (C) Patients with protein non-stable mutation had shortest median DFS than patients with protein stable mutation and TP53 wild-type patients. (D) Patients with protein non-stable mutation had shortest median OS than patients with protein stable mutation and TP53 wild-type patients.

Notes: Protein stable mutations would include non-truncating and non-frame altering mutations outside of the p53 tetramerization domain, and protein non-stable mutations would include all truncating and frame-altering mutations, as well as mutations in the tetramerization domain.

Figure 5 Survival analyses by Kaplan–Meier according to TP53 mutation type in the DNA binding domain. (A and B) Patients with non-missense mutations in the DNA binding domain had a significantly shorter median DFS and OS than TP53 wild-type patients and those with missense mutations in the DNA binding domain.

Table 2 Univariate and Multivariate Cox Regression Analysis of DFS in TP53 Wild-Type and -Mutated Patients

	TP53 Wild-Type Group							TP53 Mutated Group
	Univariate					Multivariate		Univariate					Multivariate
	N	%	DFS			HR (95% CI)	p value	N	%	DFS			HR (95% CI)	p value
			mDFS (Range, Month)	HR (95% CI)	p value					mDFS (Range, Month)	HR (95% CI)	p value
Age at diagnosis (years)
≤50	64	59.3	51.0(9.0–197.0)	1.00		1.00		49	62.0	28.0(4.0–148.0)	1.00		1.00
>50	44	40.7	50.0(16.0–197.0)	1.09(0.70–1.70)	0.713	1.22(0.55–2.67)	0.626	30	38.0	40.0(7.0–131.0)	0.77(0.44–1.36)	0.363	0.44(0.20–0.98)	0.044
Family history
No	84	77.8	52.0(9.0–197.0)	1.00		1.00		62	78.5	32.0(5.0–148.0)	1.00		1.00
Breast/ovarian cancer	7	6.5	30.0(10.0–124.0)	1.62(0.65–4.05)	0.300	1.20(0.14–10.20)	0.870	3	3.8	23.0(19.0–123.0)	0.75(0.23–2.46)	0.634	3.24(0.41–25.81)	0.267
Other cancers	17	15.7	46.5(9.0–125.0)	1.28(0.69–2.39)	0.435	1.52(0.43–5.32)	0.515	14	17.7	39.0(4.0–73.0)	1.12(0.54–2.32)	0.756	0.84(0.37–1.94)	0.688
Stage at diagnosis
I–II	52	56.5	49.5(9.0–177.0)	1.00		1.00		29	40.8	21.5(4.0–72.0)	1.00		1.00
III	22	23.9	36.0(9.0–197.0)	1.11(0.66–1.87)	0.703	2.20(1.08–4.48)	0.030	22	31.0	28.0(5.0–148.0)	0.70(0.39–1.27)	0.241	0.78(0.38–1.58)	0.487
IV	18	19.6	NA	NA	NA	NA	NA	20	28.2	NA	NA	NA	NA	NA
Grade
I–II	53	80.3	49.0(9.0–177.0)	1.00		1.00		41	75.9	32.0(9.0–148.0)	1.00		1.00
III	13	19.7	41.0(17.0–197.0)	0.69(0.34–1.40)	0.310	1.04(0.20–5.49)	0.966	13	24.1	21.5(4.0–89.0)	1.52(0.77–3.01)	0.227	0.70(0.12–4.17)	0.699
Ki67
≤20%	35	38.9	60.5(16.0–177.0)	1.00		1.00		35	47.9	43.0(7.0–148.0)	1.00		1.00
>20%	55	61.1	34.0(9.0–133.0)	2.40(1.46–3.96)	0.001	1.90(0.99–3.64)	0.054	38	52.1	21.0(4.0–131.0)	1.96(1.10–3.49)	0.022	3.47(1.53–7.89)	0.003
HR status
Positive	85	78.7	56.0(9.0–197.0)	1.00		1.00		53	67.1	46.0(4.0–148.0)	1.00		1.00
Negative	23	21.3	26.0(9.0–197.0)	1.58(0.94–2.66)	0.087	2.09(0.97–4.49)	0.059	26	32.9	19.0(5.0–51.0)	4.31(2.24–8.27)	0.000	4.40(1.97–9.84)	0.000
HER2 status
Positive	15	13.9	34.0(17.0–71.0)	1.00		1.00		21	26.6	21.0(9.0–131.0)	1.00		1.00
Negative	93	86.1	52.0(9.0–197.0)	0.45(0.23–0.90)	0.024	0.48(0.19–1.20)	0.117	58	73.4	33.0(4.0–148.0)	0.85(0.45–1.59)	0.600	1.18(0.49–2.83)	0.710
Lymph node status
Negative	42	39.3	51.0(9.0–197.0)	1.00		1.00		34	43.6	31.0(4.0–131.0)	1.00		1.00
Positive	65	60.7	51.0(14.0–197.0)	1.10(0.71–1.70)	0.663	0.98(0.23–4.22)	0.976	44	56.4	39.0(5.0–148.0)	0.85(0.50–1.44)	0.538	1.00(0.28–3.55)	1.00
Disease involvement
Non-visceral	42	39.6	40.5(9.0–153.0)	1.00		1.00		32	40.5	29.5(5.0–148.0)	1.00		1.00
Visceral	64	60.4	52.0(14.0–197.0)	0.70(0.45–1.10)	0.124	1.43(0.68–3.01)	0.343	46	59.5	36.0(4.0–123.0)	0.87(0.49–1.54)	0.640	1.65(0.74–3.69)	0.222
Adjuvant endocrine therapy
TAM/TOR	41	67.2	52.0(9.0–197.0)	1.00		1.00		23	62.2	41.5(4.0–148.0)	1.00		1.00
AI	14	23.0	53.0(16.0–177.0)	0.92(0.49–1.73)	0.792	0.52(0.13–2.06)	0.354	10	27.0	28.5(7.0–131.0)	1.30(0.61–2.78)	0.501	1.53(0.43–5.42)	0.508
TAM plus AI	3	4.9	100.5(63.0–138.0)	0.50(0.12–2.10)	0.343	1.00(0.12–8.68)	0.998	3	8.1	50.0(32.0–72.0)	0.95(0.28–3.21)	0.932	1.05(0.09–12.54)	0.967
Others	3	4.9	153.0(35.0–197.0)	0.32(0.10–1.09)	0.069	NA	NA	1	2.7	NA	0.36(0.05–2.78)	0.330	NA	NA
Adjuvant chemotherapy
PTX	9	12.5	60.0(23.0–122.0)	1.00		1.00		12	25.0	39.0(23.0–123.0)	1.00		1.00
Anthracycline	23	31.9	55.5(18.0–197.0)	0.77(0.35–1.69)	0.507	3.29(0.78–13.89)	0.106	7	14.6	34.0(5.0–66.0)	1.72(0.66–4.48)	0.265	1.38(0.18–10.32)	0.754
PTX plus anthracycline	36	50.0	48.0(9.0–177.0)	1.03(0.49–2.18)	0.932	0.78(0.24–2.59)	0.688	25	52.1	24.0(9.0–131.0)	1.43(0.71–2.88)	0.323	1.59(0.35–7.22)	0.550
Others	4	5.6	44.0(30.0–84.0)	1.30(0.40–4.25)	0.661	2.26(0.19–27.58)	0.523	4	8.3	72.0(9.0–117.0)	0.73(0.20–2.63)	0.629	0.47(0.02–12.22)	0.650

Abbreviations: HER2, human epidermal growth factor receptor 2; ER, estrogen receptor; PR, progesterone receptor; mDFS, median disease free survival; HR, hazard ratio; CI, confidence interval; NA, unknow; AI, aromatase inhibitor; TAM, tamoxifen; TOR, toremifene; PTX, paclitaxel.

Table 3 Clinical Characteristics of Patients Receiving Adjuvant Endocrine Therapy (n=96)

Characteristics	TP53 Status		p value
	Wild-Type (n=59)	Mutated (n=37)
Age at diagnosis (years)
≤50	38(64.4%)	26(70.3%)	0.553
>50	21(35.6%)	11(29.7%)
Stage at diagnosis
I~II	36(61.0%)	19(51.4%)	0.273
III~IV	16(27.1%)	14(37.8%)
Unknow	7(11.9%)	4(10.8%)
Grade
I–II	30(50.8%)	14(37.8%)	0.257
III	7(11.9%)	1(2.7%)
Unknow	22(37.3%)	22(59.5%)
Ki67
1~20%	24(40.7%)	15(40.5%)	0.820
>20%	26(44.1%)	18(48.7%)
Unknow	9(15.2%)	4(10.8%)
Lymph node status
Positive	37(62.7%)	22(59.5%)	0.928
Negative	21(35.6%)	13(35.1%)
Unknow	1(1.7%)	2(5.4%)
Disease involvement
Visceral	38(64.4%)	23(62.2%)	0.824
Non-visceral	21(35.6%)	14(37.8%)
Adjuvant endocrine therapy
Tamoxifen/toremifene	41(69.5%)	23(62.2%)	0.238
Aromatase inhibitor	14(23.7%)	10(27.0%)
Tamoxifen+ aromatase inhibitor	3(5.1%)	3(8.1%)
Others	1(1.7%)	1(2.7%)
Adjuvant chemotherapy
Yes	50(84.7%)	29(78.4%)	0.467
No	8(13.6%)	7(18.9%)
Unknow	1(1.7%)	1(2.7%)
ESR1 mutation
With ESR1 mutation	7(11.9%)	3(8.1%)	0.558
Without ESR1 mutation	52(88.1%)	34(91.9%)

Figure 6 Survival analyses by Kaplan–Meier according to TP53 status in MBC receiving adjuvant endocrine therapy. (A) There was no significant difference in TP53 status in the endocrine therapy-resistant cohort. (B) TP53 wild-type patients had a significantly better clinical outcome than TP53-mutated patients in the endocrine therapy sensitive cohort.