Christoforos Haritos - Academia.edu (original) (raw)

Papers by Christoforos Haritos

Journal for ImmunoTherapy of Cancer, Apr 18, 2017

Background: Tumor immune cell infiltrates are essential in hindering cancer progression and may c... more Background: Tumor immune cell infiltrates are essential in hindering cancer progression and may complement the TNM classification. CD8+ and CD163+ cells have prognostic impact in breast cancer but their spatial heterogeneity has not been extensively explored in this type of cancer. Here, their potential as prognostic biomarkers was evaluated, depending on their combined densities in the tumor center (TC) and the tumor invasive margin (IM). Methods: CD8+ and CD163+ cells were quantified by immunohistochemistry of formalin-fixed, paraffinembedded (FFPE) tumor tissue samples from a cohort totaling 162 patients with histologically-confirmed primary invasive non-metastatic ductal breast cancer diagnosed between 2000 and 2015. Clinical follow-up (median 6.9 years) was available for 97 of these patients. Results: Differential densities of CD8+ and CD163+ cells in the combined TC and IM compartments (i.e., high(H)/ low(L), respectively for CD8+ cells and the reverse L/H combination for CD163+ cells) were found to have significant prognostic value for survival, and allowed better patient stratification than TNM stage, tumor size, lymph node invasion and histological grade. The combined evaluation of CD8+ and CD163+ cell densities jointly in TC and IM further improves prediction of clinical outcomes based on disease-free and overall survival. Patients having the favorable immune signatures had favorable clinical outcomes despite poor clinicopathological parameters. Conclusions: Given the important roles of CD8+ and CD163+ cells in regulating opposing immune circuits, adding an assessment of their differential densities to the prognostic biomarker armamentarium in breast cancer would be valuable. Larger validation studies are necessary to confirm these findings.

Breast Cancer Research and Treatment, Oct 27, 2016

Purpose Breast cancer is a leading cause of cancer deaths in women, but despite steady improvemen... more Purpose Breast cancer is a leading cause of cancer deaths in women, but despite steady improvements in therapies, treatment is still suboptimal. Immunotherapy holds promise as a more effective therapy for breast cancer; supporting this, our prior study showed that patients possessing HER2-reactive CD8? T cells in blood experience survival superior to patients without these cells. Here, we define a composite set of biomarkers that identify patients with T cell responses to tumour antigens. Methods We assessed T cell responses following in vitro stimulation with the HER2, MUC1 and SUR tumour-associated antigens (TAA) by flow cytometry and intracellular cytokine staining in 50 breast cancer patients. We also measured HLA type, serum cytokines, tumour-infiltrating leukocytes and blood leukocyte populations. Results We found few correlations between TAA-reactive T cells and HLA type, serum cytokines and tumour-infiltrating leukocytes, whereas blood leukocyte phenotypes broadly correlated with TAA responses. This showed monocytes, natural killer cells, dendritic cells and T cells to be inversely associated with both CD4? and CD8? T cells reactive to tumour antigens. Moreover, combining multiple parameters improved the accuracy in predicting patients with TAA-responsive T cells. Conclusion This study therefore defines composite immune profiles that identify patients responding to TAAs which may allow better personalisation of cancer therapies. Keywords Tumour-associated antigen Á Blood leukocytes Á Breast cancer Á HER2 Á Survivin Á MUC1 Abbreviations AJCC American Joint Committee on Cancer ER Oestrogen receptor FoxP3 Forkhead box P3 HER2 Human epidermal growth factor receptor 2 HLA Human leukocyte antigen Nicole Janssen and Sotirios P. Fortis have contributed equally to this work.

Journal for ImmunoTherapy of Cancer, 2015

Journal for ImmunoTherapy of Cancer, May 30, 2017

After publication of this article [1], it was noticed that Table 3 contained errors introduced du... more After publication of this article [1], it was noticed that Table 3 contained errors introduced during the production process. The corrected Table 3 can be seen below and the original article has been updated to correct the errors.

Journal for ImmunoTherapy of Cancer, Nov 4, 2015

Clinical and Experimental Medicine, Feb 12, 2018

Novel molecular markers that address the heterogeneity of breast cancer (BC) and provide meaningf... more Novel molecular markers that address the heterogeneity of breast cancer (BC) and provide meaningful prognostic information for BC patients are needed. Kallikrein-related peptidase 6 (KLK6) is aberrantly expressed and functionally implicated in BC and, like other members of the KLK family, may prove a useful molecular tool for clinical management. Our objective was to assess, for the first time, the clinical relevance of KLK6 mRNA expression in BC. Total RNA was isolated from 165 breast tumors, as well as 100 adjacent non-cancerous tumor specimens. After cDNA synthesis, and following quality control, quantitative real-time PCR for KLK6 expression analysis took place. Receiver operating characteristic curves were constructed in order to assess the ability of KLK6 mRNA expression levels to differentiate between molecular BC subtypes. Survival analyses, using DFS as endpoint, were performed at the univariate and multivariate levels. Publicly available BC databases and online survival analysis tools were used to validate our findings. A significant downregulation of KLK6 mRNA expression was observed in BC tissue sections compared to the non-cancerous component (P < 0.001). The expression of KLK6 is positively associated with tumor grade (P = 0.038) and is overexpressed in TNBC and HER2-positive tumors (P < 0.001). Aberrant KLK6 expression predicts the clinical outcome of BC patients in terms of DFS, independently of currently used prognostic markers (HR = 7.11, 95% CI = 1.19-42.45). The differential expression of KLK6 and its association with unfavorable outcome in BC patients was validated via in silico analyses. Although an independent external cohort is necessary to confirm our findings, we proved for the first time that KLK6 can provide independent prognostic information for BC patients.

Οι όγκοι του μαστού παρουσιάζουν εκτενή ετερογένεια η οποία αντανακλάται από ποικίλα βιολογικά κα... more Οι όγκοι του μαστού παρουσιάζουν εκτενή ετερογένεια η οποία αντανακλάται από ποικίλα βιολογικά και μοριακά χαρακτηριστικά, από την προγνωστική έκβαση και την απόκριση στη θεραπεία. Νέοι μοριακοί δείκτες που παρέχουν σημαντική προγνωστική πληροφορία για τις ασθενείς με καρκίνο του μαστού είναι απαραίτητοι. Σύμφωνα με προγενέστερες μελέτες, η έκφραση των μελών της οικογένειας των καλλικρεϊνών συχνά συσχετίζεται με την κλινική έκβαση των ασθενών με καρκίνο του μαστού. Επιπλέον, η Kallikrein-related peptidase 6 (KLK6-Καλλικρεΐνη 6) έχει αναφερθεί ότι εμπλέκεται στον καρκίνο του μαστού και ως εκ τούτου δύναται να αποδειχθεί ένα χρήσιμο μοριακό εργαλείο για κλινική χρήση.Ολικό RNA εξήχθη από 165 όγκους μαστού και από 100 παρακείμενους μη καρκινικούς ιστούς. Μετά τη σύνθεση συμπληρωματικού DNA (cDNA) και τον επακόλουθο ποιοτικό έλεγχο, τα επίπεδα του mRNA αναλύθηκαν με τη χρήση ποσοτικής σε πραγματικό χρόνο αλυσιδωτής αντίδρασης πολυμεράσης (real time PCR). Χαρακτηριστικές λειτουργικές καμ...

DFS and OS in patients according to standard clinicopathological variables. Figures S2-S5. DFS (A... more DFS and OS in patients according to standard clinicopathological variables. Figures S2-S5. DFS (A, C) and OS (B, D) for patients stratified by grade (Figure S2), T status (Figure S3), nodal status (Figure S4) and pathological TNM stage (Figure S5) and analyzed according to the density of CD8+ (A, B) or CD163+ (C, D) cells in TC or IM. Figures S6-S9. Kaplan-Meier curves illustrating DFS (A, C, E) and OS (B, D, E) for patients stratified by grade (Figure S6), T status (Figure S7), nodal status (Figure S8) and pathological TNM stage (Figure S9) and analyzed according to the density of CD8+ (A, B) or CD163+ (C, D) cells in the combined tumor regions and to the combined immune signatures (E, F). The statistics with significant differences or strong trends between groups and the corresponding hazard ratios are shown in the respective plots. (PDF 356Â kb)

Journal for immunotherapy of cancer, 2017

After publication of this article [1], it was noticed that Table 3 contained errors introduced du... more After publication of this article [1], it was noticed that Table 3 contained errors introduced during the production process. The corrected Table 3 can be seen below and the original article has been updated to correct the errors.

Journal for immunotherapy of cancer, Jan 18, 2017

Tumor immune cell infiltrates are essential in hindering cancer progression and may complement th... more Tumor immune cell infiltrates are essential in hindering cancer progression and may complement the TNM classification. CD8+ and CD163+ cells have prognostic impact in breast cancer but their spatial heterogeneity has not been extensively explored in this type of cancer. Here, their potential as prognostic biomarkers was evaluated, depending on their combined densities in the tumor center (TC) and the tumor invasive margin (IM). CD8+ and CD163+ cells were quantified by immunohistochemistry of formalin-fixed, paraffin-embedded (FFPE) tumor tissue samples from a cohort totaling 162 patients with histologically-confirmed primary invasive non-metastatic ductal breast cancer diagnosed between 2000 and 2015. Clinical follow-up (median 6.9 years) was available for 97 of these patients. Differential densities of CD8+ and CD163+ cells in the combined TC and IM compartments (i.e., high(H)/low(L), respectively for CD8+ cells and the reverse L/H combination for CD163+ cells) were found to have s...

Breast cancer research and treatment, 2017

Breast cancer is a leading cause of cancer deaths in women, but despite steady improvements in th... more Breast cancer is a leading cause of cancer deaths in women, but despite steady improvements in therapies, treatment is still suboptimal. Immunotherapy holds promise as a more effective therapy for breast cancer; supporting this, our prior study showed that patients possessing HER2-reactive CD8+ T cells in blood experience survival superior to patients without these cells. Here, we define a composite set of biomarkers that identify patients with T cell responses to tumour antigens. We assessed T cell responses following in vitro stimulation with the HER2, MUC1 and SUR tumour-associated antigens (TAA) by flow cytometry and intracellular cytokine staining in 50 breast cancer patients. We also measured HLA type, serum cytokines, tumour-infiltrating leukocytes and blood leukocyte populations. We found few correlations between TAA-reactive T cells and HLA type, serum cytokines and tumour-infiltrating leukocytes, whereas blood leukocyte phenotypes broadly correlated with TAA responses. Thi...

Journal for ImmunoTherapy of Cancer, 2015

Journal for ImmunoTherapy of Cancer, 2015

Journal for ImmunoTherapy of Cancer, Apr 18, 2017

Background: Tumor immune cell infiltrates are essential in hindering cancer progression and may c... more Background: Tumor immune cell infiltrates are essential in hindering cancer progression and may complement the TNM classification. CD8+ and CD163+ cells have prognostic impact in breast cancer but their spatial heterogeneity has not been extensively explored in this type of cancer. Here, their potential as prognostic biomarkers was evaluated, depending on their combined densities in the tumor center (TC) and the tumor invasive margin (IM). Methods: CD8+ and CD163+ cells were quantified by immunohistochemistry of formalin-fixed, paraffinembedded (FFPE) tumor tissue samples from a cohort totaling 162 patients with histologically-confirmed primary invasive non-metastatic ductal breast cancer diagnosed between 2000 and 2015. Clinical follow-up (median 6.9 years) was available for 97 of these patients. Results: Differential densities of CD8+ and CD163+ cells in the combined TC and IM compartments (i.e., high(H)/ low(L), respectively for CD8+ cells and the reverse L/H combination for CD163+ cells) were found to have significant prognostic value for survival, and allowed better patient stratification than TNM stage, tumor size, lymph node invasion and histological grade. The combined evaluation of CD8+ and CD163+ cell densities jointly in TC and IM further improves prediction of clinical outcomes based on disease-free and overall survival. Patients having the favorable immune signatures had favorable clinical outcomes despite poor clinicopathological parameters. Conclusions: Given the important roles of CD8+ and CD163+ cells in regulating opposing immune circuits, adding an assessment of their differential densities to the prognostic biomarker armamentarium in breast cancer would be valuable. Larger validation studies are necessary to confirm these findings.

Breast Cancer Research and Treatment, Oct 27, 2016

Purpose Breast cancer is a leading cause of cancer deaths in women, but despite steady improvemen... more Purpose Breast cancer is a leading cause of cancer deaths in women, but despite steady improvements in therapies, treatment is still suboptimal. Immunotherapy holds promise as a more effective therapy for breast cancer; supporting this, our prior study showed that patients possessing HER2-reactive CD8? T cells in blood experience survival superior to patients without these cells. Here, we define a composite set of biomarkers that identify patients with T cell responses to tumour antigens. Methods We assessed T cell responses following in vitro stimulation with the HER2, MUC1 and SUR tumour-associated antigens (TAA) by flow cytometry and intracellular cytokine staining in 50 breast cancer patients. We also measured HLA type, serum cytokines, tumour-infiltrating leukocytes and blood leukocyte populations. Results We found few correlations between TAA-reactive T cells and HLA type, serum cytokines and tumour-infiltrating leukocytes, whereas blood leukocyte phenotypes broadly correlated with TAA responses. This showed monocytes, natural killer cells, dendritic cells and T cells to be inversely associated with both CD4? and CD8? T cells reactive to tumour antigens. Moreover, combining multiple parameters improved the accuracy in predicting patients with TAA-responsive T cells. Conclusion This study therefore defines composite immune profiles that identify patients responding to TAAs which may allow better personalisation of cancer therapies. Keywords Tumour-associated antigen Á Blood leukocytes Á Breast cancer Á HER2 Á Survivin Á MUC1 Abbreviations AJCC American Joint Committee on Cancer ER Oestrogen receptor FoxP3 Forkhead box P3 HER2 Human epidermal growth factor receptor 2 HLA Human leukocyte antigen Nicole Janssen and Sotirios P. Fortis have contributed equally to this work.

Journal for ImmunoTherapy of Cancer, 2015

Journal for ImmunoTherapy of Cancer, May 30, 2017

After publication of this article [1], it was noticed that Table 3 contained errors introduced du... more After publication of this article [1], it was noticed that Table 3 contained errors introduced during the production process. The corrected Table 3 can be seen below and the original article has been updated to correct the errors.

Journal for ImmunoTherapy of Cancer, Nov 4, 2015

Clinical and Experimental Medicine, Feb 12, 2018

Novel molecular markers that address the heterogeneity of breast cancer (BC) and provide meaningf... more Novel molecular markers that address the heterogeneity of breast cancer (BC) and provide meaningful prognostic information for BC patients are needed. Kallikrein-related peptidase 6 (KLK6) is aberrantly expressed and functionally implicated in BC and, like other members of the KLK family, may prove a useful molecular tool for clinical management. Our objective was to assess, for the first time, the clinical relevance of KLK6 mRNA expression in BC. Total RNA was isolated from 165 breast tumors, as well as 100 adjacent non-cancerous tumor specimens. After cDNA synthesis, and following quality control, quantitative real-time PCR for KLK6 expression analysis took place. Receiver operating characteristic curves were constructed in order to assess the ability of KLK6 mRNA expression levels to differentiate between molecular BC subtypes. Survival analyses, using DFS as endpoint, were performed at the univariate and multivariate levels. Publicly available BC databases and online survival analysis tools were used to validate our findings. A significant downregulation of KLK6 mRNA expression was observed in BC tissue sections compared to the non-cancerous component (P < 0.001). The expression of KLK6 is positively associated with tumor grade (P = 0.038) and is overexpressed in TNBC and HER2-positive tumors (P < 0.001). Aberrant KLK6 expression predicts the clinical outcome of BC patients in terms of DFS, independently of currently used prognostic markers (HR = 7.11, 95% CI = 1.19-42.45). The differential expression of KLK6 and its association with unfavorable outcome in BC patients was validated via in silico analyses. Although an independent external cohort is necessary to confirm our findings, we proved for the first time that KLK6 can provide independent prognostic information for BC patients.

Οι όγκοι του μαστού παρουσιάζουν εκτενή ετερογένεια η οποία αντανακλάται από ποικίλα βιολογικά κα... more Οι όγκοι του μαστού παρουσιάζουν εκτενή ετερογένεια η οποία αντανακλάται από ποικίλα βιολογικά και μοριακά χαρακτηριστικά, από την προγνωστική έκβαση και την απόκριση στη θεραπεία. Νέοι μοριακοί δείκτες που παρέχουν σημαντική προγνωστική πληροφορία για τις ασθενείς με καρκίνο του μαστού είναι απαραίτητοι. Σύμφωνα με προγενέστερες μελέτες, η έκφραση των μελών της οικογένειας των καλλικρεϊνών συχνά συσχετίζεται με την κλινική έκβαση των ασθενών με καρκίνο του μαστού. Επιπλέον, η Kallikrein-related peptidase 6 (KLK6-Καλλικρεΐνη 6) έχει αναφερθεί ότι εμπλέκεται στον καρκίνο του μαστού και ως εκ τούτου δύναται να αποδειχθεί ένα χρήσιμο μοριακό εργαλείο για κλινική χρήση.Ολικό RNA εξήχθη από 165 όγκους μαστού και από 100 παρακείμενους μη καρκινικούς ιστούς. Μετά τη σύνθεση συμπληρωματικού DNA (cDNA) και τον επακόλουθο ποιοτικό έλεγχο, τα επίπεδα του mRNA αναλύθηκαν με τη χρήση ποσοτικής σε πραγματικό χρόνο αλυσιδωτής αντίδρασης πολυμεράσης (real time PCR). Χαρακτηριστικές λειτουργικές καμ...

DFS and OS in patients according to standard clinicopathological variables. Figures S2-S5. DFS (A... more DFS and OS in patients according to standard clinicopathological variables. Figures S2-S5. DFS (A, C) and OS (B, D) for patients stratified by grade (Figure S2), T status (Figure S3), nodal status (Figure S4) and pathological TNM stage (Figure S5) and analyzed according to the density of CD8+ (A, B) or CD163+ (C, D) cells in TC or IM. Figures S6-S9. Kaplan-Meier curves illustrating DFS (A, C, E) and OS (B, D, E) for patients stratified by grade (Figure S6), T status (Figure S7), nodal status (Figure S8) and pathological TNM stage (Figure S9) and analyzed according to the density of CD8+ (A, B) or CD163+ (C, D) cells in the combined tumor regions and to the combined immune signatures (E, F). The statistics with significant differences or strong trends between groups and the corresponding hazard ratios are shown in the respective plots. (PDF 356Â kb)

Journal for immunotherapy of cancer, 2017

After publication of this article [1], it was noticed that Table 3 contained errors introduced du... more After publication of this article [1], it was noticed that Table 3 contained errors introduced during the production process. The corrected Table 3 can be seen below and the original article has been updated to correct the errors.

Journal for immunotherapy of cancer, Jan 18, 2017

Tumor immune cell infiltrates are essential in hindering cancer progression and may complement th... more Tumor immune cell infiltrates are essential in hindering cancer progression and may complement the TNM classification. CD8+ and CD163+ cells have prognostic impact in breast cancer but their spatial heterogeneity has not been extensively explored in this type of cancer. Here, their potential as prognostic biomarkers was evaluated, depending on their combined densities in the tumor center (TC) and the tumor invasive margin (IM). CD8+ and CD163+ cells were quantified by immunohistochemistry of formalin-fixed, paraffin-embedded (FFPE) tumor tissue samples from a cohort totaling 162 patients with histologically-confirmed primary invasive non-metastatic ductal breast cancer diagnosed between 2000 and 2015. Clinical follow-up (median 6.9 years) was available for 97 of these patients. Differential densities of CD8+ and CD163+ cells in the combined TC and IM compartments (i.e., high(H)/low(L), respectively for CD8+ cells and the reverse L/H combination for CD163+ cells) were found to have s...

Breast cancer research and treatment, 2017

Breast cancer is a leading cause of cancer deaths in women, but despite steady improvements in th... more Breast cancer is a leading cause of cancer deaths in women, but despite steady improvements in therapies, treatment is still suboptimal. Immunotherapy holds promise as a more effective therapy for breast cancer; supporting this, our prior study showed that patients possessing HER2-reactive CD8+ T cells in blood experience survival superior to patients without these cells. Here, we define a composite set of biomarkers that identify patients with T cell responses to tumour antigens. We assessed T cell responses following in vitro stimulation with the HER2, MUC1 and SUR tumour-associated antigens (TAA) by flow cytometry and intracellular cytokine staining in 50 breast cancer patients. We also measured HLA type, serum cytokines, tumour-infiltrating leukocytes and blood leukocyte populations. We found few correlations between TAA-reactive T cells and HLA type, serum cytokines and tumour-infiltrating leukocytes, whereas blood leukocyte phenotypes broadly correlated with TAA responses. Thi...

Journal for ImmunoTherapy of Cancer, 2015

Journal for ImmunoTherapy of Cancer, 2015