Association of TRF2 expression and myeloid-derived suppressor cells infiltration with clinical outcome of patients with cutaneous melanoma

Figures & data

Table 1. Clinical and histomolecular characteristics of the metastatic melanoma cohorts treated by chemotherapy, targeted therapy, or immunotherapy. *χ2-test or Student’s t-test were used to investigate difference between groups.

Characteristics	Patients treated by chemotherapy (n = 54), 43.2%	Patients treated by immunotherapy (n = 48), 38.4%	Patients treated by targeted therapy (n = 23), 18.4%	Total (n = 125), 100%	p-value*	Test
Gender					0.21	χ2-test
Female	22 (40.7%)	14 (29.2%)	5 (21.7%)	41 (32.8%)
Male	32 (59.3%)	34 (70.8%)	18 (78.3%)	84 (67.2%)
Age (years)					0.18	ANOVA
Mean	66.7	63	60.6	64.2
Range	[23–92]	[24–89]	[26–87]	[23–92]
ECOG status					0.84	Fisher’s test
0	41 (75.9%)	38 (79.2%)	18 (78.3%)	97 (77.6%)
1	6 (11.1%)	5 (10.4%)	3 (13%)	14 (11.2%)
2	5 (9.3%)	5 (10.4%)	1 (4.3%)	11 (8.8%)
3	2 (3.7%)	0 (0%)	1 (4.3%)	3 (2.4%)
LDH baseline					0.45	Fisher’s test
Normal	26 (48.1%)	23 (47.9%)	16 (69.6%)	65 (52%)
High	5 (9.3%)	2 (4.2%)	4 (17.4%)	11 (8.8%)
Not determined	23 (42.6%)	23 (47.9%)	3 (13%)	49 (39.2%)
Histological subtype					<0.001	Fisher’s test
Superficial spreading melanoma	26 (48.1%)	15 (31.2%)	15 (65.2%)	56 (44.8%)
Nodular melanoma	19 (35.2%)	10 (20.8%)	3 (13%)	32 (25.6%)
Acral lentiginous melanoma	3 (5.6%)	3 (6.2%)	0 (0%)	6 (4.8%)
Invasive lentigo maligna melanoma	3 (5.6%)	1 (2.1%)	1 (4.3%)	5 (4%)
Not classified	3 (5.6%)	19 (39.6%)	4 (17.4%)	26 (20.8%)
Ulceration					0.84	χ2-test
Absent	25 (46.3%)	24 (50%)	11 (47.8%)	60 (48%)
Present	29 (53.7%)	22 (45.8%)	12 (52.2%)	63 (50.4%)
Unknown	0 (0%)	2 (4.2%)	0 (0%)	2 (1.6%)
Stage at diagnosis					0.01	χ2-test
I + II	47 (87%)	32 (66.7%)	21 (91.3%)	100 (80%)
III + IV	7 (13%)	16 (33.3%)	2 (8.7%)	25 (20%)
Breslow depth (median, range)					0.71	Kruskal-Wallis test
Median, range	2.5 [0.15–12]	3.1 [0.3–25]	3 [0.22–10]	2.6 [0.15–25]
Brain metastasis (at diagnosis)					0.11	χ2-test
Present	3 (5.6%)	6 (12.5%)	5 (21.7%)	14 (11.2%)
Absent	51 (94.4%)	42 (87.5%)	18 (78.3%)	111 (88.8%)
BRAF status					<0.001	χ2-test
Mutation	19 (35.2%)	3 (6.3%)	18 (78.3%)	40 (32%)
Wild-type	35 (64.8%)	28 (58.3%)	2 (8.7%)	65 (52%)
Unknown	0 (0%)	17 (35.4%)	3 (13%)	20 (16%)
BRAF mutation type					0.78	Fisher’s test
p.V600E	13/19 (68.4%)	3/3 (100%)	13/18 (72.2%)	29/40 (72.5)
p.V600K	5/19 (26.3%)	0/3 (0%)	3/18 (16.7%)	8/40 (20)
p.V600D	0/19 (0%)	0/3 (0%)	1/18 (5.6%)	1/40 (2.5%)
p.V600R	0/19 (0%)	0/3 (0%)	1/18 (5.6%)	1/40 (2.5%)
p.L597R	1/19 (5.3%)	0/3 (0%)	0/18 (0%)	1/40 (2.5%)

Figure 1. Representative images of immune biomarkers and TRF2 staining, and their cell detection mask overlays used in the digital image analysis. Original magnification, x 200

Table 2. Correlative analysis between the clinical and histomolecular characteristics of the patients and the analyzed biomarkers in the metastatic melanoma cohort. *χ2-test, Student’s t-test or ANOVA test were used to investigate difference between groups.

Characteristics	CD14			CD15			CD33			TRF2
	Negative (n = 63), 50.4%	Positive (n = 62), 49.6%	P-value*	Negative (n = 63), 50.4%	Positive (n = 62), 49.6%	P-value*	Negative (n = 63), 50.4%	Positive (n = 62), 49.6%	P-value*	Negative (n = 63), 50.4%	Positive (n = 62), 49.6%	P-value*
Gender			0.66			0.75			0.66			0.95
Female	19 (30.2%)	22 (35.5%)		22 (34.9%)	19 (30.6%)		19 (30.2%)	22 (35.5%)		20 (31.7%)	21 (33.9%)
Male	44 (69.8%)	40 (64.5%)		41 (65.1%)	43 (69.4%)		44 (69.8%)	40 (64.5%)		43 (68.3%)	41 (66.1%)
Age (years)			0.73			0.14			0.16			0.63
Mean	64.6	63.7		62.3	66.1		66	62.3		64.8	63.6
Range	[23–88]	[26–92]		[23–88]	[24–92]		[26–92]	[23–89]		[24–89]	[23–92]
ECOG status			0.73			0.23			0.22			0.38
0	49 (77.8%)	48 (77.4%)		51 (81%)	46 (74.2%)		45 (71.4%)	52 (83.9%)		46 (73%)	51 (82.3%)
1	6 (9.5%)	8 (12.9%)		8 (12.7%)	6 (9.7%)		9 (14.3%)	5 (8.1%)		10 (15.9%)	4 (6.5%)
2	7 (11.1%)	4 (6.5%)		4 (6.3%)	7 (11.3%)		8 (12.7%)	3 (4.8%)		6 (9.5%)	5 (8.1%)
3	1 (1.6%)	2 (3.2%)		0 (0%)	3 (4.8%)		1 (1.6%)	2 (3.2%)		1 (1.6%)	2 (3.2%)
LDH baseline			0.85			1			1			0.75
Normal	30 (47.6%)	35 (56.5%)		34 (54%)	31 (50%)		36 (57.1%)	29 (46.8%)		29 (46%)	36 (58.1%)
High	6 (9.5%)	5 (8.1%)		6 (9.5%)	5 (8.1%)		6 (9.5%)	5 (8.1%)		4 (6.3%)	7 (11.3%)
Not determined	27 (42.9%)	22 (35.5%)		23 (36.5%)	26 (41.9%)		21 (33.3%)	28 (45.2%)		30 (47.6%)	19 (30.6%)
Histological subtype			0.12			0.39			0.08			< 0.001
Superficial spreading melanoma	24 (38.1%)	32 (51.6%)		31 (49.2%)	25 (40.3%)		26 (41.3%)	30 (48.4%)		18 (28.6%)	38 (61.3%)
Nodular melanoma	21 (33.3%)	11 (17.7%)		17 (27%)	15 (24.2%)		22 (34.9%)	10 (16.1%)		13 (20.6%)	19 (30.6%)
Acral lentiginous melanoma	1 (1.6%)	5 (8.1%)		1 (1.6%)	5 (8.1%)		1 (1.6%)	5 (8.1%)		3 (4.8%)	3 (4.8%)
Invasive lentigo maligna melanoma	3 (4.8%)	2 (3.2%)		3 (4.8%)	2 (3.2%)		3 (4.8%)	2 (3.2%)		4 (6.3%)	1 (1.6%)
Not classified	14 (22.2%)	12 (19.4%)		11 (17.5%)	15 (24.2%)		11 (17.5%)	15 (24.2%)		25 (39.7%)	1 (1.6%)
Ulceration			1			0.005			0.24			0.79
Absent	30 (47.6%)	30 (48.4%)		39 (61.9%)	21 (33.9%)		27 (42.9%)	33 (53.2%)		31 (49.2%)	29 (46.8%)
Present	32 (50.8%)	31 (50%)		24 (38.1%)	39 (62.9%)		36 (57.1%)	27 (43.5%)		30 (47.6%)	33 (53.2%)
Not determined	1 (1.6%)	1 (1.6%)		0 (0%)	2 (3.2%)		0 (0%)	2 (3.2%)		2 (3.2%)	0 (0%)
Stage at diagnosis			0.69			0.35			0.08			0.2
I + II	49 (77.8%)	51 (82.3%)		53 (84.1%)	47 (75.8%)		46 (73%)	54 (87.1%)		47 (74.6%)	53 (85.5%)
III + IV	14 (22.2%)	11 (17.7%)		10 (15.9%)	15 (24.2%)		17 (27%)	8 (12.9%)		16 (25.4%)	9 (14.5%)
Breslow depth (median)			0.62			0.02			0.06			0.49
Median	2.6	3		2.2	3.3		3.6	2.4		3.5	2.5
Range	[0.15–12]	[0.2–25]		[0.15–25]	[0.95–25]		[0.15–25]	[0.39–10]		[0.15–12]	[0.2–25]
Brain metastasis (at diagnosis)			1			0.8			0.4			0.8
Present	7 (11.1%)	7 (11.3%)		8 (12.7%)	6 (9.7%)		9 (14.3%)	5 (8.1%)		8 (12.7%)	6 (9.7%)
Absent	56 (88.9%)	55 (88.7%)		55 (87.3%)	56 (90.3%)		54 (85.7%)	57 (91.9%)		55 (87.3%)	56 (90.3%)
BRAF status			0.02			0.56			0.52			0.72
Mutation	14 (22.2%)	26 (41.9%)		19 (30.2%)	21 (33.9%)		20 (31.7%)	20 (32.3%)		15 (23.8%)	25 (40.3%)
Wild-type	40 (63.5%)	25 (40.3%)		36 (57.1%)	29 (46.8%)		38 (60.3%)	27 (43.5%)		28 (44.4%)	37 (59.7%)
Not determined	9 (14.3%)	11 (17.7%)		8 (12.7%)	12 (19.4%)		5 (7.9%)	15 (24.2%)		20 (31.7%)	0 (0%)
Treatment type			0.37			0.62			0.72			0.004
Anti-BRAF	3 (4.8%)	9 (14.5%)		4 (6.3%)	8 (12.9%)		7 (11.1%)	5 (8.1%)		5 (7.9%)	7 (11.3%)
Anti-BRAF + anti-MEK	2 (3.2%)	4 (6.5%)		2 (3.2%)	4 (6.5%)		4 (6.3%)	2 (3.2%)		1 (1.6%)	5 (8.1%)
Anti-MEK	0 (0%)	1 (1.6%)		0 (0%)	1 (1.6%)		0 (0%)	1 (1.6%)		0 (0%)	1 (1.6%)
Chemotherapy	26 (41.3%)	23 (37.1%)		26 (41.3%)	23 (37.1%)		25 (39.7%)	24 (38.7%)		17 (27%)	32 (51.6%)
Ipilimumab	9 (14.3%)	5 (8.1%)		8 (12.7%)	6 (9.7%)		7 (11.1%)	7 (11.3%)		10 (15.9%)	4 (6.5%)
Ipilimumab + Nivolumab	5 (7.9%)	3 (4.8%)		3 (4.8%)	5 (8.1%)		2 (3.2%)	6 (9.7%)		6 (9.5%)	2 (3.2%)
Nivolumab	18 (28.6%)	17 (27.4%)		20 (31.7%)	15 (24.2%)		18 (28.6%)	17 (27.4%)		24 (38.1%)	11 (17.7%)

Figure 2. Kaplan-Meier overall survival curves according to TFR2, CD33, and CD33+/CD15+/CD14- status in the whole study population (n = 125)

Figure 3. Kaplan-Meier overall survival curve according to CD33+/CD15+/CD14- status in patients treated by immunotherapy (n = 48)

Table 3. Multivariate analysis for overall survival in the cohort population

Variables	Sub-groups	HR multivariate (95% CI; P-value)
ECOG status	0	-
	≥ 1	1.86 (1.07–3.23; P=.03)
Brain metastasis at diagnosis	Absent	-
	Present	1.07 (0.54–2.14; P =.85)
Type of treatment	Chemotherapy	-
	Immunotherapy	0.95 (0.54–1.67; P =.85)
	Targeted therapy	0.97 (0.53–1.78; P =.93)
Breslow depth	Mean (SD)	0.97 (0.91–1.02; P =.25)
Ulceration	Absent	-
	Present	0.87 (0.55–1.39; P =.57)
Stage at diagnosis	I–II	-
	III–IV	1.25 (0.70–2.24; P=.44)
CD33	Low density/mm^2	-
	High density/mm^2	1.12 (0.7–2.1; P=.097)
TRF2	Low density/mm^2	-
	High density/mm^2	1.14 (1.1–1.46; P=.179)
CD33+/CD15+/CD14-	Low density/mm^2	-
	High density/mm^2	2.6 (1.1–4.6; P=.037)