The effect of CD39+CD8+ T cells and CD39 and CD73 on tumor cells on the treatment response of epithelial ovarian cancer and patient outcomes

doi:10.21203/rs.3.rs-2366378/v1

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The effect of CD39+CD8+ T cells and CD39 and CD73 on tumor cells on the treatment response of epithelial ovarian cancer and patient outcomes

https://doi.org/10.21203/rs.3.rs-2366378/v1

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Objective. In this study, we evaluated whether CD39+CD8+ tumor-infiltrating lymphocytes (TILs) and CD39 and CD73 on tumor cells had an effect on survival in epithelial ovarian cancer (EOC) patients treated with platinum-based chemotherapy.

Methods. A total of 129 EOC patients between 2009 and 2015 were retrospectively reviewed. The expression of CD39+CD8+ TILs in the TME and CD39 and CD73 on tumor cells was measured by multiplexed quantitative fluorescence (QIF) on tissue microarrays (TMAs).

Results. The expression of CD39+CD8+ TILs in the TME and CD39 and CD73 on tumor cells in EOC patients was negatively associated with the response to platinum-based treatment. The area under the ROC curve of CD39+CD8+ TILs and CD39 and CD73 on tumor cells IHC image quantification for predicting the platinum resistance in EOC patients was 0.679, 0.718, 0.682; respectively. Furthermore, the protein expression of CD39+CD8+ TILs and CD39 and CD73 on tumor cells were higher in the platinum Resistant group than in the Sensitive group (P<0.01) and associated with a worse prognosis. Kaplan-Meier plots and Cox model analysis demonstrated that overexpression of CD39+CD8+ TILs and CD39 and CD73 on tumor cells was associated with a significantly shorter PFS (HR: 1.76, 95% CI: 1.20-2.56; HR: 1.64, 95% CI: 1.10-2.42; HR: 1.47, 95% CI: 0.99-2.16, respectively) and decreased OS (HR: 1.66, 95% CI: 1.12-2.47; HR: 1.78, 95% CI: 1.19-2.66; HR: 1.52, 95% CI: 1.02-2.27, respectively) in EOC patients.

Conclusion. Our results show that the expression of CD39+CD8+ TILs and CD39 and CD73 on tumor cells could be useful markers of treatment response and prognosis in EOC patients treated with platinum-based chemotherapy in Chinese women.

Epithelial ovarian cancer

CD39+CD8+ T cell

CD39

CD73

Platinum-based chemotherapy

Prognosis

Epithelial ovarian cancer (EOC) is the third most common gynecologic malignancy in the world, but it has the highest mortality rate among these cancers [1]. Due to a lack of effective screening strategies and the absence of symptoms in the early stages of the disease, most cases of EOC are already at an advanced stage in the initial diagnosis [2]. Despite improved clinical detection methods and therapies, the 5-year survival rate for EOC patients with advanced-stage disease has not changed over the past several decades [3], suggesting that a better understanding of the progression and metastasis mechanisms of EOC might contribute to changing the course of this disease and improving survival.

Tumor progression is dependent not only on the characteristics of malignant cells but also on the behavior of the entire tumor microenvironment (TME). High levels of CD8 + tumor-infiltrating lymphocytes (TILs) in the TME can improve the survival of patients with a wide variety of tumor types, including ovarian cancer [4, 5]. However, TILs are a highly heterogeneous cell population across patients and tumors, and it is difficult to identify tumor-specific CD8⁺ lymphocytes. Evan et al. [6] found that CD39 was highly expressed by tumor-specific CD8 + TILs and that the frequencies of CD39 + CD8 + T cells correlated with clinical parameters in lung cancer. In contrast, bystander CD8 + TILs strikingly lack CD39 expression, suggesting that CD39 + CD8 + T cells include more tumor reactive T cells.

CD39, a transmembrane extracellular ATPase, is widely expressed by activated lymphocytes, regulatory T cells, B cells and some tumor cells [7]. In conjunction with CD73, CD39 can degrade extracellular ATP and ADP to adenosine, which is an important tumor-induced immunosuppressive mechanism [8, 9]. Accumulated evidence also shows that the overexpression of CD39 and CD73 on tumor cells is associated with tumor invasiveness and metastasis [10, 11] and with shorter patient survival time [12], indicating that the CD39/CD73 adenosine pathway is closely involved in cancer progression.

It is unclear whether these immune response-related factors are related to the clinical prognosis of EOC. In this study, we investigated the association between the expression levels of CD39 + CD8 + TILs in the TME and CD39 and CD73 on tumor cells and the response to chemotherapy and prognosis of EOC patients.

Tissue Samples Collection

The study included tissue specimens of 129 patients diagnosed with primary EOC treated between October 2009 and November 2015 at the Department of Gynecology, the Fourth Hospital of Hebei Medical University. All participants were regularly followed up for 5 years. The study was approved by the Ethics Committee of the Fourth Hospital of Hebei Medical University (NO. 2020K-1164), and informed consent was obtained from all of the recruited subjects according to the Helsinki Declaration.

All patients received platinum-based chemotherapy after primary debulking surgery for six to eight cycles. The chemotherapeutic regimen consisted of paclitaxel (175 mg/m²), carboplatin (area under curve 5) or cisplatin (75 mg/m²). Progression-free survival (PFS) and overall survival (OS) denote the time interval from the date of ovarian carcinoma surgery to the events of tumor recurrence and death regardless of cause or date of last contact, respectively [13]. The platinum-free interval (PFI) was defined as the interval between the completion of platinum-based chemotherapy infusion and the date of disease progression [14]. According to the PFI, all patients were stratified into three groups: from < 6 months, < 6 to 12 months vs. more than 12 months. Platinum-sensitive group (Sensitive group) and platinum-resistant group (Resistant group) were defined as previously described [15]. Patients with a PFI more than 12 months were categorized as platinum-sensitive, whereas patients with a PFI < 6 months and < 6 to 12 months were categorized as platinum-resistant and partially platinum-sensitive, respectively.

The eligibility inclusion criteria included newly diagnosed, histologically confirmed EOC patients who received platinum-based chemotherapy after primary debulking surgery. In contrast, patients were excluded if neoadjuvant chemotherapy was administered and no other primary neoplasm was present.

Establishment Of Tissue Microarrays (Tmas)

149 formalin-fixed paraffin-embedded (FFPE) EOC samples were used to establish TMAs. 16 due to FFPE tissues missing, and 4 patients lost to follow-up, the laboratory data were analyzed in 129 patients. FFPE tissue blocks of all cases were designated by 2 independent pathologists, and highly representative areas of invasive carcinoma were marked on the H&E stained slides. Using these H&E slides for reference, 0.6 mm core biopsies were obtained from each tumor specimen using a needle and arrayed on a recipient paraffin block (Beecher Instruments, Silver Spring, MD, USA. Figure 1).

Immunohistochemistry (Ihc) Assay

An IHC assay was utilized to detect the CD8, CD39 and CD73 levels in the TMA sections. Briefly, 4-µm-thick FFPE tissue sections were treated at room temperature for 15 min with 3% hydrogen peroxide-containing methanol to block the endogenous peroxidase activity. After washing, the slides were incubated with the primary antibodies as follows: rabbit anti-CD8 (ab217344, Abcam, 1:2000), rabbit anti-CD39 (ab223842, Abcam, 1:1000), and rabbit anti-CD73 (D7F9A, Cell Signaling, 1:200). Subsequently, using a Dako autostainer with the EnVision™ FLEX/HRP + detection system (EnVision™ FLEX DAB kit), and then hematoxylin was applied for staining. The CD8, CD39 and CD73 expression was analyzed based on the number of positive cells per mm² by the Vectra System (PerkinElmer, Waltham, MA, USA). The median of the positive cells per mm² was a watershed between positive or negative individuals.

Multiplexed Immunofluorescence Assay

Slides of TMA were heated in a 63°C oven for 1h, deparaffinized and rehydrated in xylene and a series of graded alcohols (100%, 95%, 75% and 50%) before pH 8 EDTA antigen retrieval. For nonspecific antigen blocking, slides were immersed in a blocking solution containing 0.3% bovine serum albumin in 0.05% Tween solution for 30 minutes. After washing, slides were then incubated with the following primary antibodies in order: anti-CD8 (ab217344, Abcam, 1:2000), anti-CD39 (ab223842, Abcam, 1:1000), anti-CD73 (D7F9A, Cell Signaling, 1:200), anti-pancytokeratin (PA125, Baidao Medical Technology). The secondary antibody (Dako, Glostrup, Denmark) was then added and incubated. Staining with Opal dye reagents (diluted 1:100 and incubated for 6 min) was repeated 5 times. Each cycle consisted of blocking, incubation with primary antibody and secondary antibody, Opal dye, and removal of surface-bound antibody. Sections were counter-stained with DAPI to identify all nuclei. H&E staining and immunofluorescence pictures were captured using a Vectra Polaris Automated Quantitative Pathology Imaging System (Perkin Elmer, Waltham, MA).

Multiplexed Quantitative Immunofluorescence (Qif)

Quantitative measurement of fluorescent protein expression was performed using the automated quantitative analysis (AQUA) method, as described previously [16]. Fields of view (FOV) were selected that were cytokeratin positive or adjacent to a cytokeratin positive FOV in a previously acquired low resolution image. A total compartment consisting of cytokeratin positivity (e.g., epithelial regions), absence of cytokeratin positivity (e.g., stromal regions), or DAPI-positive cells (e.g., total tissue including epithelial and stroma) was generated by the DAPI signal and cytokeratin signal, respectively. For analysis with TissueFAXS software (TissueGnostics), the QIF scores of each fluorescence channel were calculated by dividing the target TIL marker pixel intensity within each compartment by the area. Stained slides were examined by two pathologists, and only cases with more than three cytokeratin-positive FOVs were included.

Kaplan-meier Plotter Database Analysis

The prognostic value of CD8, CD39 and CD73 mRNA levels in patients with advanced EOC was determined using the “K-M plotter” database, integrated gene expression data and survival information of over 1,000 advanced EOC patients. In this study, PFS and OS of EOC patients for 5 years were evaluated using the K-M plotter. The exclude outlier arrays were selected as the array quality control.

Statistical Analysis

The data were analyzed using SPSS version 23.0 software (SPSS Inc., Chicago, IL, USA). Using the survival ROC package in R software version 3.5.0, QIF scores were split into low and high populations at a cutoff value objectively determined by ROC curve analysis. The chi-square test and Fisher’s exact test were used to compare CD39 + CD8 + TILs in the TME and CD39 and CD73 on tumor cells and clinical pathological variables of patients. ROC curve analysis was employed for calculating the diagnostic value of CD39 + CD8 + TILs in the TME and CD39 and CD73 on tumor cells on platinum resistance. Survival curves were plotted using the Kaplan‒Meier method, and comparisons among groups were performed using log-rank tests. A Cox proportional hazards ratio model was used for multivariate analysis. All P values were based on two-sided tests, and p < 0.05 was considered statistically significant.

Demographic Profile

Baseline demographic and clinical characteristics are shown in Table 1. Of the 129 EOC patients included in this study, the median age was 62 years (range, 33–89 years). In terms of FIGO staging, 119 cases (92.2%) were in stage III and 10 cases (7.8%) in stages IV. According to pathology type, there were 88 cases (68.2%) of high-grade serous ovarian cancer (HGSOC), 33 (25.6%) of endometrioid carcinoma, and 8 (6.2%) of other types.

Table 1

Baseline characteristics of patients with advanced epithelial ovarian cancer.
Characteristics	All patients (%)
Age
< 60	77 (59.7)
≥ 60	52 (40.3)
FIGO stage
III	119 (92.2)
IV	10 (7.8)
Pathology
HGSOC	88 (68.2)
Endometrioid	33 (25.6)
Others	8 (6.2)
Tumor residual size
0cm	29 (22.5)
≥ 1cm	38 (29.5)
> 1cm	62 (48.1)
Platinum-based regimen
TC	90 (69.8)
TP	32 (24.8)
Other	7 (5.4)
PFI
< 6 mo	16 (12.4)
< 6–12 mo	37 (28.7)
≥ 12 mo	92 (71.3)
HGSOC, high grade serous ovarian cancer.

Platinum-based Chemotherapy Response

Ninety-two (71.3%, PFI > 12 months) patients responded to first-line therapy (platinum-sensitive), and 37 (28.7%, including PFI < 6 months and PFI < 6–12 months) patients were platinum-resistant. By the immunofluorescence assay of the tissue microarray, the relationship between the expression levels of CD39 + CD8 + TILs, CD39 and CD73 on tumor cells and platinum resistance was further explored. The levels of CD39 + CD8 + TILs, CD39 and CD73 on tumor cells were significantly different between the two groups (Table 2).

Table 2

Association of CD39 + CD8 + TIL, CD39 and CD73 on tumor cells protein expression with chemo-response.
	Expression	Resistant group	Sensitive group	P	OR	95%CI
CD39 + CD8 + TIL	low	10 (17.9)	46 (82.1)	0.019	2.700	1.174–6.209
CD39 + CD8 + TIL	high	27 (37.0)	46 (63.0)	0.019	2.700	1.174–6.209
CD39	low	20 (50.0)	20 (50.0)	0.001	4.235	1.876–9.564
CD39	high	72 (80.9)	17 (19.1)	0.001	4.235	1.876–9.564
CD73	low	25 (56.8)	19 (43.2)	0.010	2.829	1.282–6.243
CD73	high	67 (78.8)	18 (21.2)	0.010	2.829	1.282–6.243

The expression of CD39 + CD8 + TILs, CD39 and CD73 on tumor cells was related to platinum resistance

There were significant differences in the platinum-based chemotherapy response of cases between different expression of CD39 + CD8 + TILs (P < 0.0001; Fig. 2A). Patients with high levels of CD39 + CD8 + TILs had a decreased response to platinum-based chemotherapy compared to those with low levels (OR: 2.70, 95% CI: 1.17–6.21). To investigate the value of CD39 + CD8 + TILs in platinum resistance in EOC, ROC analysis demonstrated that the area under the curve (AUC) was 0.679 (95%CI: 0.570–0.787) and the cut-off value was 73; with sensitivity = 0.659; 1-specificity = 0.875; Youden’s index = 0.334) (Fig. 2D).

Consistently, patients with high levels of CD39 on tumor cells had a decreased response to platinum-based chemotherapy compared to those with low levels (P < 0.0001; OR: 4.24, 95% CI: 1.88–9.56; Fig. 2B). ROC analysis demonstrated that the AUC was 0.718 (95%CI: 0.622–0.814) and the cut-off value was 243.5; with sensitivity = 0.692; 1-specificity = 0.615; Youden’s index = 0.308) (Fig. 2E).

Similarly, patients with high levels of CD73 on tumor cells had a decreased response to platinum-based chemotherapy compared to those with low levels (P < 0.0001; OR: 2.83, 95% CI: 1.28–6.24; Fig. 2C). ROC analysis demonstrated that the AUC was 0.682 (95%CI: 0.585–0.779) and the cut-off value was 70.5; with sensitivity = 0.696; 1-specificity = 0.693; Youden’s index = 0.389) (Fig. 2F).

These findings suggested that high expression of CD39 + CD8 + TILs, CD39 and CD73 on tumor cells was associated with platinum resistance.

Survival

Platinum resistance is one major reason of the high mortality in EOC patients

In the present study, the Kaplan-Meier survival analysis was utilized to compare PFS and OS curves between the Resistant group and the Sensitive group. The results showed that patients in the chemotherapy Resistant group had a shortened PFS and OS than the Sensitive group (P = 0.035 and P = 0.028, respectively) (Figs. 3A, B). In a 5-year follow-up study, the median PFS of EOC patients in the Resistant group and the Sensitive group were 16 and 24 months, respectively, and the median OS of those patients were 29 and 44 months, respectively.

High Expression Of Cd39 + cd8 + tils Was Associated With Poor Prognosis In Eoc Patients

As illustrated in Fig. 4A, the expression levels of CD39 + CD8 + TILs in TMAs were identified. The median PFS of patients with high and low levels of CD39 + CD8 + TILs was 15 and 22 months, respectively, and the median OS of those patients was 22 and 28, respectively. Compared with patients with low levels of CD39 + CD8 + TILs, patients with high levels of CD39 + CD8 + TILs had a shorter median PFS and median OS time. Kaplan-Meier plots illustrate the differences in PFS and OS for patients with low and high levels of CD39 + CD8 + TILs (Fig. 4B, C). Kaplan-Meier plots and Cox model analysis showed that the levels of CD39 + CD8 + TILs were associated with PFS (HR = 1.76, 95% CI = 1.20–2.56) and OS (HR = 1.66, 95% CI = 1.12–2.47; Table 3) of EOC patients.

Table 3

Association of CD39 + CD8 + TIL, CD39 and CD73 on tumor cells with prognosis.
	Recurrence		P	HR	95%CI	Survival		P	HR	95%CI
	Yes (n %)	No (n %)	P	HR	95%CI	No (n %)	Yes (n %)	P	HR	95%CI
CD39 + CD8 + TIL
low	45(80.4)	11(19.6)	0.003	1.755	1.202–2.563	41(73.2)	15(26.8)	0.010	1.664	1.120–2.474
high	68(93.2)	5(6.8)	0.003	1.755	1.202–2.563	62(84.9)	11(15.1)	0.010	1.664	1.120–2.474
CD39
low	75(84.3)	14(15.7)	0.011	1.635	1.104–2.421	66(74.2)	23(25.8)	0.004	1.777	1.188–2.660
high	38(95.0)	2(5.0)	0.011	1.635	1.104–2.421	37(92.5)	3(7.5)	0.004	1.777	1.188–2.660
CD73
low	72(84.7)	13(15.3)	0.043	1.468	0.998–2.158	63(74.1)	22(25.9)	0.043	1.522	1.021–2.269
high	41(93.2)	3(6.8)	0.043	1.468	0.998–2.158	40(90.9)	4(9.1)	0.043	1.522	1.021–2.269

Furthermore, K-M Plotter database [http://kmplot.com/analysis] was used for prognostic analysis. Elevated mRNA expression of CD39 + CD8 + TILs was associated with significantly shorter PFS (HR = 1.22, 95%CI = 1.05–1.41; P = 0.0076) (Fig. 4D) for all EOC patients, and OS in grade III, IV EOC patients (HR = 1.20, 95% CI = 1.02–1.41; P = 0.03) (Fig. 4E) followed for 5 years. The median PFS of EOC patients with CD39 + CD8 + TILs mRNA high expression and low expression was 18.83 and 24.09 months, respectively. The median OS of these patients was 38.73 and 44.00 months, respectively.

High Expression Of Cd39 On Tumor Cells Was Associated With Poor Prognosis In Eoc Patients

As presented in Fig. 5A, we identified the expression levels of CD39 on tumor cells in TMAs of EOC patients. The Kaplan-Meier survival analysis demonstrated the significant differences in PFS (p = 0.011, Fig. 5B) and OS (P = 0.004, Fig. 5C) for patients with low and high levels of CD39 on tumor cells. Compared with patients with low levels of CD39 on tumor cells, patients with high levels of CD39 on tumor cells had a shorter median PFS and median OS time. The median PFS of patients with high and low levels of CD39 on tumor cells was 11 and 20 months, respectively, and the median OS of those patients was 25 and 37, respectively. Kaplan-Meier plots and Cox model analysis showed that the levels of CD39 on tumor cells were associated with PFS (HR = 1.64, 95% CI = 1.10–2.42) and OS (HR = 1.78, 95% CI = 1.19–2.66; Table 3) of EOC patients.

Using the “K-M plotter” database, high mRNA expression of CD39 on tumor cells was associated with significantly shorter PFS (HR = 1.29, 95%CI = 1.11–1.5; P = 0.00071) (Fig. 5D) in stage III, IV EOC patients, and OS for advanced serous patients (HR = 1.22, 95% CI = 1.01–1.48; P = 0.041) (Fig. 5E) followed for 5 years. The median PFS of EOC patients with CD39 on tumor cells mRNA high expression and low expression was 13.37 and 18 months, respectively. The median OS of these patients was 37.93 and 43 months, respectively.

High Expression Of Cd73 On Tumor Cells Was Associated With Poor Prognosis In Eoc Patients

Representative images of the expression levels of CD73 on tumor cells of TMAs were illustrated in Fig. 6A. The Kaplan-Meier plots demonstrated marked differences in PFS (p = 0.043, Fig. 6B) and OS (P = 0.043, Fig. 6C) for patients with low and high levels of CD73 on tumor cells. Compared with patients with low levels of CD73 on tumor cells, patients with high levels of CD73 on tumor cells had a shorter mean OS time. The median PFS of patients with high and low levels of CD73 on tumor cells was 13 and 20 months, respectively, and the mean OS of those patients was 33 and 37, respectively. Kaplan-Meier plots and Cox model analysis showed that the levels of CD73 on tumor cells were associated with the survival prognosis PFS (HR = 1.47, 95% CI = 1.00-2.16) and OS (HR = 1.52, 95% CI = 1.02–2.27; Table 3) of EOC patients.

In the present study, we used the KM plotter database and accessed the prognostic roles of CD73 on tumor cells. High mRNA expression of CD73 on tumor cells was associated with significantly shorter PFS (HR = 1.24, 95%CI = 1.06–1.46; P = 0.0082) (Fig. 6D) and OS (HR = 1.23, 95% CI = 1.03–1.46; P = 0.02) (Fig. 6E) in stage III, IV EOC patients followed for 5 years. The median PFS of EOC patients with CD73 on tumor cells mRNA high expression and low expression was 13 and 17.43 months, respectively. The median OS of these patients was 37.03 and 43.97 months, respectively.

Collectively, high expression of CD39 + CD8 + TILs, CD39 and CD73 on tumor cells may be closely associated with poor prognosis in EOC patients.

The standard initial treatment for advanced-stage EOC comprises cytoreductive surgery followed by platinum-based chemotherapy [17]. However, the recurrence rate is high, and the 5-year survival rate is only 29% in advanced EOC [3, 18]. Increasing evidence supports the TME as a fertile soil for tumor growth, resulting in tumor progression and metastasis [19]. However, the TME characteristics of EOC and its clinical significance are not well understood. We therefore investigated the effect of immune response-related factors on the treatment response, PFS and OS in EOC patients. Our results suggested that the overexpression of CD39 + CD8 + T cells and CD39 and CD73 on tumor cells decreased the platinum-based chemotherapy response, thus worsening EOC patients’ survival rate.

CD8 + T cells are functionally heterogeneous [20] and can be transferred from a functional state to a dysfunctional state [21]. In some studies, CD39 is considered to be a biomarker of exhausted T cells, although it is a marker of tumor-specific T cells [22]. Recently, the efficacy of CD39 + CD8 + T cells has been suggested to be associated with survival in patients with many solid tumors, such as esophageal squamous cell carcinoma [23], glioblastoma [24], clear cell renal cell carcinoma [25], and colorectal, head and neck and pancreatic cancers [26]. High CD39 + CD8 + T cells in the TME were associated with poor OS in the above reports. Similar findings were obtained in the present study. EOC patients with high-level expression of CD39 + CD8 + T cells had a significantly lower response to therapy and poor prognosis. By clonal expansions of CD8 + CD39 + T cells, Yost et al. demonstrated that CD8⁺CD39⁺ T cells coexpressed markers of chronic T-cell activation and exhaustion and were evident in patients with basal or squamous cell carcinoma [27]. The immunological characteristics of CD39 + CD8 + TILs from both the primary tumor site and metastatic sites exhibit severe T-cell exhaustion features in EOC [28]. Accordingly, increased CD8⁺CD39⁺ T cells in the TME may lead to downregulation of the immune response against tumors [25].

In the TME, CD39 is expressed by infiltrating lymphocytes, the tumor stroma, and tumor cells. CD39 in cancer cells displays ATPase activity and, together with CD73, generates adenosine [29]. Overexpression of CD39 and CD73 has been associated with a poorer prognosis for several cancers [30–33]. In human triple-negative breast cancer, high expression of CD73 promotes resistance to anthracycline and is a poor prognosis factor [34]. However, the treatment response in ovarian cancer has rarely been reported. In this study, our results revealed that higher levels of CD39 and CD73 on tumor cells were associated with a significantly lower response to platinum-based chemotherapy, thus worsening survival in EOC patients. Using ovarian cancer cell lines, Hausler and colleagues [35] showed that ovarian cancer cell lines express CD39 and produce adenosine, which suppresses T-cell–mediated antitumor responses, further strengthening our conclusions. Moreover, the presence of CD39 and CD73 in ovarian cancer produces adenosine, which can be 30–60 times higher than that produced by Tregs and augments the immunosuppressive effect of adenosine on the host immune system [36]. Although the mechanisms of the association between increased CD39 and CD73 and a poorer treatment response have not been fully clarified, the level of CD39 and CD73 on cancer cells may reflect the balance between extracellular ATP, ADP, AMP and adenosine, which is decisive in the control of tumor progression [37].

In conclusion, our results demonstrate that the overexpression of CD39 + CD8 + T cells and CD39 and CD73 on tumor cells was associated with a poor chemo-response and a reduced survival of EOC patients. Novel therapies are needed to treat this type of cancer. More importantly, the adenosine pathway, including CD73, CD39, and adenosine receptors, has emerged as a key metabolic pathway that regulates tumor immunity [38–40], indicating that monoclonal antibodies targeting CD73 or CD39 could be more promising and efficient tools for cancer therapy. According to our data, EOC patients with overexpression of CD39 + CD8 + T cells and CD39 and CD73 on tumor cells have a poor prognosis; simultaneously, however, they are also more likely to benefit from immunotherapy or a combination with chemotherapy. To our knowledge, this is the first report demonstrating that overexpression of CD39 + CD8 + T cells and CD39 and CD73 on tumor cells had a decreased response to platinum-based chemotherapy. Follow-up studies are needed to strengthen these conclusions.

Ethics Approval and Consent to Participate

The study was approved by the Ethics Committee of the Fourth Hospital of Hebei Medical University (NO. 2020K-1164), and informed consent was obtained from all of the recruited subjects according to the Helsinki Declaration.

Consent for Publication

Not applicable.

Competing Interests

The authors declare that they have no conflict of interest.

Funding

This study was supported by the Natural Science Foundation of Hebei Province [grant number: H2020206385]; and the Scientific Research Fund of Hebei Provincial Health and Family Planning Commission [grant number: 20200099].

Author Contributions

Yan Li, Shan Kang, and Hai-Yan Sun designed the study and carried out the experiments. Hai-Yan Sun, Jian-Lei Wu and Xue-Ping Li recruited the patients and collected the data. Hai-Yan Sun, Yue-Ping Liu and Yan Li analyzed the data and prepared draft figures and tables. All authors were involved in writing the paper and provided final approval of the submitted and published versions.

Acknowledgements

We thank the patients, their families, and the investigators who participated in this study.

Availability of Data and Material

The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.

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The effect of CD39+CD8+ T cells and CD39 and CD73 on tumor cells on the treatment response of epithelial ovarian cancer and patient outcomes

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Abstract

Figures

Introduction

Materials And Methods

Tissue Samples Collection

Establishment Of Tissue Microarrays (Tmas)

Immunohistochemistry (Ihc) Assay

Multiplexed Immunofluorescence Assay

Multiplexed Quantitative Immunofluorescence (Qif)

Kaplan-meier Plotter Database Analysis

Statistical Analysis

Results

Demographic Profile

Platinum-based Chemotherapy Response

Survival

Platinum resistance is one major reason of the high mortality in EOC patients

High Expression Of Cd39 + cd8 + tils Was Associated With Poor Prognosis In Eoc Patients

High Expression Of Cd39 On Tumor Cells Was Associated With Poor Prognosis In Eoc Patients

High Expression Of Cd73 On Tumor Cells Was Associated With Poor Prognosis In Eoc Patients

Discussion

Declarations

References

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