A type I interferon footprint in pre-operative biopsies is an independent biomarker that in combination with CD8⁺ T cell quantification can improve the prediction of response to neoadjuvant treatment of rectal adenocarcinoma

Figures & data

Table 1. Demographic characteristics of the study population.

Patients n		130
Female n (%)		43 (33)
Age mean (range)		63 (25–88)
Clinical stage n (%)	T1–2	7 (5)
	T3	62 (48)
	T4	59 (45)
	TX	2 (2)
	N0	35 (27)
	N1–2	93 (72)
	NX	2 (2)
	M0	111 (85)
	M1	19 (15)
Pathological stage n (%)	T0	8 (6)
	T1	3 (2)
	T2	27 (21)
	T3	68 (52)
	T4	19 (15)
	missing	5 (4)
	N0	59 (45)
	N1	42 (32)
	N2	24 (18)
	missing	5 (4)
UICC stage n (%)	0	11 (8)
	1	13 (10)
	2	31 (24)
	3	54 (42)
	4	19 (15)
	missing	2 (2)
Neoadjuvant tr. n (%)	SCRT	57 (44)
	CRT	68 (52)
	CT	5 (4)
Surgical procedure n (%)	Ant. Res.	41 (32)
	Hartmann	14 (11)
	APE	73 (56)
	Other	2 (2)
TRG (AJCC) n (%)	0	13 (10)
	1	16 (12)
	2	58 (45)
	3	43 (33)

SCRT: Short course Radiotherapy (5×5 Gy), CRT: Chemoradiotherapy, CT: Chemotherapy, Ant.Res.: Anterior Resection.

Figure 1. Enumeration of T cells subsets in tumor tissue according to the effectiveness of treatment. (a) Bar charts show the mean density ±SEM of the CD3.⁺ T cells, CD3⁺CD8⁺ T cells, CD3⁺CD8⁺GrzB⁺ Cytotoxic T cells, and γδTCR⁺cells per mm²of the area according to the patients’ response to treatment in whole tumor tissue. (b) In the epithelium 50µm surrounding area (c) and only epithelium. (d) Bar charts show the mean density ±SEM of the T cells subsets per mm² of the whole tumor tissue in complete responders v.s non-responders in whole tumor tissue. (e) In the epithelium and the area with 50µm width around it (f) and only epithelium. GrzB: Granzyme B, TRG: Tumor Regression Grade. Wilcoxon's rank sum test p <0.05 considered significant; n=130.

Figure 2. Enumeration of MxA⁺ cells in tumor stroma. (a) Bar charts show the mean density ±SEM of MxA⁺ Pan-CK⁻ cells per mm² of the tumor stroma according to the patients’ response to the treatment. (b) Bar charts show the mean density ±SEM of the MxA⁺ cells per mm² of tumor stroma amongst complete responders and non-responders. TRG: Tumor Regression Grade. Wilcoxon's rank sum test; p <0.05 considered significant; n=130.

Figure 3. Classification of the patients according to the effectiveness of treatment. (a)The frequency of patients with CD3⁺ (Left-Upper), CD8⁺ (Middle –Upper) and MxA⁺ (Right-Upper), low (Blue), intermediate (White), and high (Red) according to tumor regression and the frequency of IS_B (Left-Lower) and IS_MxA+CD8⁺ (Right-Lower) low, intermediate, and high according to tumor regression. (b) The frequency of patients with either CD8⁺ or MxA⁺ high, intermediate and low according to the tumor regression. TRG: Tumor Regression Grade. Unilateral linear-by-linear association test; p <0.05 considered significant; n=130.

Table 2. Sensitivity, specificity, and diagnostic odds ratio of immune markers in prediction of response to treatment.

	Sensitivity	Specificity	Diagnostic odds ratio
CD3	0.38	0.71	6.48
CD8	0.54	0.73	16.53
MxA	0.62	0.74	27.52
ISB	0.46	0.74	10.53
ISMxA+CD8^+	0.23	0.76	2.67
MxA or CD8 (Heat-map)	0.77	0.64	83.33

Data availability statement

Data are available within the article and supplementary material.