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Myeloid derived suppressor and dendritic cell subsets are related to clinical outcome in prostate cancer patients treated with prostate GVAX and ipilimumab.

Santegoets SJ, Stam AG, Lougheed SM, Gall H, Jooss K, Sacks N, Hege K, Lowy I, Scheper RJ, Gerritsen WR, van den Eertwegh AJ, de Gruijl TD - J Immunother Cancer (2014)

Bottom Line: Redressing this balance may therefore be of clinical benefit.Significant treatment-induced activation of conventional and plasmacytoid DC subsets (cDC and pDC) was observed, which in the case of BDCA1/CD1c(+) cDC1 and MDC8(+)/6-sulfoLacNAc(+) inflammatory cDC3 was associated with significantly prolonged overall survival (OS), but also with the development of autoimmune-related adverse events.High pre-treatment levels of CD14(+)HLA-DR(-)monocytic MDSC (mMDSC) were associated with reduced OS.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.

ABSTRACT

Background: Cancer-related disturbances in myeloid lineage development, marked by high levels of myeloid-derived suppressor cells (MDSC) and impaired dendritic cell (DC) development, are associated with poor clinical outcome due to immune escape and therapy resistance. Redressing this balance may therefore be of clinical benefit. Here we investigated the effects of combined Prostate GVAX/ipilimumab immunotherapy on myeloid subsets in peripheral blood of castration-resistant prostate cancer (CRPC) patients as well as the putative predictive value of baseline and on-treatment myeloid parameters on clinical outcome.

Methods: Patients with CRPC (n = 28) received thirteen intradermal administrations of Prostate GVAX, consisting of two allogeneic GM-CSF-transduced and irradiated prostate cancer cell lines (LN-CaP and PC3) and six infusions of escalating doses of anti-CTLA4/ipilimumab. Frequencies and activation status of peripheral blood DC (PBDC) and MDSC were determined before, during and after treatment by flowcytometric analysis and related to clinical benefit.

Results: Significant treatment-induced activation of conventional and plasmacytoid DC subsets (cDC and pDC) was observed, which in the case of BDCA1/CD1c(+) cDC1 and MDC8(+)/6-sulfoLacNAc(+) inflammatory cDC3 was associated with significantly prolonged overall survival (OS), but also with the development of autoimmune-related adverse events. High pre-treatment levels of CD14(+)HLA-DR(-)monocytic MDSC (mMDSC) were associated with reduced OS. Unsupervised clustering of these myeloid biomarkers revealed particular survival advantage in a group of patients with high treatment-induced PBDC activation and low pretreatment frequencies of suppressive mMDSC in conjunction with our previously identified lymphoid biomarker of high pretreatment CD4(+)CTLA4(+) T cell frequencies.

Conclusions: Our data demonstrate that DC and MDSC subsets are affected by prostate GVAX/ipilimumab therapy and that myeloid profiling may contribute to the identification of patients with possible clinical benefit of Prostate GVAX/ipilimumab treatment.

No MeSH data available.


Related in: MedlinePlus

Prostate GVAX/ipilimumab therapy-induced activation of peripheral blood (PB) DC subsets is associated with prolonged survival. PBDC activation and frequency was determined before (week 0/visit 1 (w0v1)), during (w4v3, w8v5, w12v7, w16v9, w20v11, w24v13 and after (follow-up (fu)) prostate GVAX/ipilimumab therapy by flow cytometry. A) Activation state over treatment –by Median Fluorescence Index (MFI) of CD40 of CD11chiCD19−CD14loBDCA1+cDC1, CD11c+CD14−BDCA-3+ cDC2, CD11c+CD14loMDC8+ cDC3 and CD11c−CD14−CD123hiBDCA-2+pDC. Grey bars denote the mean ± SEM range at baseline. B) DC subset frequencies (as percentages of PBMC) and C) absolute numbers per ml blood, over treatment, cDC1: solid black squares, cDC2: open black squares, cDC3: solid grey squares, pDC: open grey squares. Means ± SEM of 28 patients are shown. D) Kaplan Meier curve for on-treatment increases in cDC1 and cDC3 activation. Number of patients and corresponding median survival for each group is given. Differences between pre- and on- or post-treatment were analyzed with the repeated measures ANOVA with a post-hoc Dunnett’s multiple comparisons test. Differences were considered significant when p < 0.05, as indicated with asterisks (* p < 0.05, ** p < 0.01) within the respective squares. Statistical significance of the survival distribution was analyzed by log-rank testing.
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Fig1: Prostate GVAX/ipilimumab therapy-induced activation of peripheral blood (PB) DC subsets is associated with prolonged survival. PBDC activation and frequency was determined before (week 0/visit 1 (w0v1)), during (w4v3, w8v5, w12v7, w16v9, w20v11, w24v13 and after (follow-up (fu)) prostate GVAX/ipilimumab therapy by flow cytometry. A) Activation state over treatment –by Median Fluorescence Index (MFI) of CD40 of CD11chiCD19−CD14loBDCA1+cDC1, CD11c+CD14−BDCA-3+ cDC2, CD11c+CD14loMDC8+ cDC3 and CD11c−CD14−CD123hiBDCA-2+pDC. Grey bars denote the mean ± SEM range at baseline. B) DC subset frequencies (as percentages of PBMC) and C) absolute numbers per ml blood, over treatment, cDC1: solid black squares, cDC2: open black squares, cDC3: solid grey squares, pDC: open grey squares. Means ± SEM of 28 patients are shown. D) Kaplan Meier curve for on-treatment increases in cDC1 and cDC3 activation. Number of patients and corresponding median survival for each group is given. Differences between pre- and on- or post-treatment were analyzed with the repeated measures ANOVA with a post-hoc Dunnett’s multiple comparisons test. Differences were considered significant when p < 0.05, as indicated with asterisks (* p < 0.05, ** p < 0.01) within the respective squares. Statistical significance of the survival distribution was analyzed by log-rank testing.

Mentions: To assess the effects of prostate GVAX/ipilimumab treatment on circulating myeloid DC subsets, frequency and activation status of circulating conventional DC (cDC) subsets cDC1, cDC2, cDC3 and plasmacytoid DC (pDC) were determined before, during and after treatment. cDC1 were identified as CD11chiCD19−CD14−BDCA-1/CD1c+; cDC2 as CD11c+CD14−BDCA-3+; cDC3 as CD11chiCD14loMDC8+ and pDC as CD11c−CD14−CD123hiBDCA-2+ (see also Additional file 2: Figure S2 for gating strategies). Similar to previous observations in cancer patients by us and by others [12,15,27,28], frequencies and activation status of circulating DC and monocytes were generally lower in CRPC patients as compared with healthy individuals (see Additional file 3: Figure S3A and 3B). On-treatment activation (shown in Figure 1A by CD40 expression levels) was observed for all DC subsets (as previously reported by us for cDC1 [11]). Interestingly, this activation was paralleled by decreases in cDC1, cDC2, and pDC frequencies (Figure 1B). These decreases were observed as early as four weeks after start of treatment and were maintained during treatment (Figure 1B). Of note, decreases in absolute cDC1, CDC2, and pDC numbers per volume blood were much less pronounced and on-treatment increased absolute cDC3 numbers even reached significance (Figure 1C). These differences between DC frequencies and absolute numbers may in large part be explained by a sustained increase in absolute lymphocyte numbers over the course of treatment (see Additional file 4: Figure S4). Increased PBDC activation was generally maintained during treatment (see Figure 1A) and increases of >70% of CD40 med. FI on the cDC1 and cDC3 subsets (see for representative histograms Additional file 2: Figure S2) were associated with significantly prolonged overall survival (OS; median survival 38.5 vs. 15.5 months, p = 0.0004 and median survival 40 vs. 19 months, p = 0.0031, respectively (Table 1). Survival benefit was even more pronounced for patients who displayed treatment-induced activation of both cDC1 and cDC3 subsets (median survival 52 vs. 16 months, p < 0.0001; Figure 1C). No relationship with survival was found for either pDC or cDC2.Figure 1


Myeloid derived suppressor and dendritic cell subsets are related to clinical outcome in prostate cancer patients treated with prostate GVAX and ipilimumab.

Santegoets SJ, Stam AG, Lougheed SM, Gall H, Jooss K, Sacks N, Hege K, Lowy I, Scheper RJ, Gerritsen WR, van den Eertwegh AJ, de Gruijl TD - J Immunother Cancer (2014)

Prostate GVAX/ipilimumab therapy-induced activation of peripheral blood (PB) DC subsets is associated with prolonged survival. PBDC activation and frequency was determined before (week 0/visit 1 (w0v1)), during (w4v3, w8v5, w12v7, w16v9, w20v11, w24v13 and after (follow-up (fu)) prostate GVAX/ipilimumab therapy by flow cytometry. A) Activation state over treatment –by Median Fluorescence Index (MFI) of CD40 of CD11chiCD19−CD14loBDCA1+cDC1, CD11c+CD14−BDCA-3+ cDC2, CD11c+CD14loMDC8+ cDC3 and CD11c−CD14−CD123hiBDCA-2+pDC. Grey bars denote the mean ± SEM range at baseline. B) DC subset frequencies (as percentages of PBMC) and C) absolute numbers per ml blood, over treatment, cDC1: solid black squares, cDC2: open black squares, cDC3: solid grey squares, pDC: open grey squares. Means ± SEM of 28 patients are shown. D) Kaplan Meier curve for on-treatment increases in cDC1 and cDC3 activation. Number of patients and corresponding median survival for each group is given. Differences between pre- and on- or post-treatment were analyzed with the repeated measures ANOVA with a post-hoc Dunnett’s multiple comparisons test. Differences were considered significant when p < 0.05, as indicated with asterisks (* p < 0.05, ** p < 0.01) within the respective squares. Statistical significance of the survival distribution was analyzed by log-rank testing.
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Fig1: Prostate GVAX/ipilimumab therapy-induced activation of peripheral blood (PB) DC subsets is associated with prolonged survival. PBDC activation and frequency was determined before (week 0/visit 1 (w0v1)), during (w4v3, w8v5, w12v7, w16v9, w20v11, w24v13 and after (follow-up (fu)) prostate GVAX/ipilimumab therapy by flow cytometry. A) Activation state over treatment –by Median Fluorescence Index (MFI) of CD40 of CD11chiCD19−CD14loBDCA1+cDC1, CD11c+CD14−BDCA-3+ cDC2, CD11c+CD14loMDC8+ cDC3 and CD11c−CD14−CD123hiBDCA-2+pDC. Grey bars denote the mean ± SEM range at baseline. B) DC subset frequencies (as percentages of PBMC) and C) absolute numbers per ml blood, over treatment, cDC1: solid black squares, cDC2: open black squares, cDC3: solid grey squares, pDC: open grey squares. Means ± SEM of 28 patients are shown. D) Kaplan Meier curve for on-treatment increases in cDC1 and cDC3 activation. Number of patients and corresponding median survival for each group is given. Differences between pre- and on- or post-treatment were analyzed with the repeated measures ANOVA with a post-hoc Dunnett’s multiple comparisons test. Differences were considered significant when p < 0.05, as indicated with asterisks (* p < 0.05, ** p < 0.01) within the respective squares. Statistical significance of the survival distribution was analyzed by log-rank testing.
Mentions: To assess the effects of prostate GVAX/ipilimumab treatment on circulating myeloid DC subsets, frequency and activation status of circulating conventional DC (cDC) subsets cDC1, cDC2, cDC3 and plasmacytoid DC (pDC) were determined before, during and after treatment. cDC1 were identified as CD11chiCD19−CD14−BDCA-1/CD1c+; cDC2 as CD11c+CD14−BDCA-3+; cDC3 as CD11chiCD14loMDC8+ and pDC as CD11c−CD14−CD123hiBDCA-2+ (see also Additional file 2: Figure S2 for gating strategies). Similar to previous observations in cancer patients by us and by others [12,15,27,28], frequencies and activation status of circulating DC and monocytes were generally lower in CRPC patients as compared with healthy individuals (see Additional file 3: Figure S3A and 3B). On-treatment activation (shown in Figure 1A by CD40 expression levels) was observed for all DC subsets (as previously reported by us for cDC1 [11]). Interestingly, this activation was paralleled by decreases in cDC1, cDC2, and pDC frequencies (Figure 1B). These decreases were observed as early as four weeks after start of treatment and were maintained during treatment (Figure 1B). Of note, decreases in absolute cDC1, CDC2, and pDC numbers per volume blood were much less pronounced and on-treatment increased absolute cDC3 numbers even reached significance (Figure 1C). These differences between DC frequencies and absolute numbers may in large part be explained by a sustained increase in absolute lymphocyte numbers over the course of treatment (see Additional file 4: Figure S4). Increased PBDC activation was generally maintained during treatment (see Figure 1A) and increases of >70% of CD40 med. FI on the cDC1 and cDC3 subsets (see for representative histograms Additional file 2: Figure S2) were associated with significantly prolonged overall survival (OS; median survival 38.5 vs. 15.5 months, p = 0.0004 and median survival 40 vs. 19 months, p = 0.0031, respectively (Table 1). Survival benefit was even more pronounced for patients who displayed treatment-induced activation of both cDC1 and cDC3 subsets (median survival 52 vs. 16 months, p < 0.0001; Figure 1C). No relationship with survival was found for either pDC or cDC2.Figure 1

Bottom Line: Redressing this balance may therefore be of clinical benefit.Significant treatment-induced activation of conventional and plasmacytoid DC subsets (cDC and pDC) was observed, which in the case of BDCA1/CD1c(+) cDC1 and MDC8(+)/6-sulfoLacNAc(+) inflammatory cDC3 was associated with significantly prolonged overall survival (OS), but also with the development of autoimmune-related adverse events.High pre-treatment levels of CD14(+)HLA-DR(-)monocytic MDSC (mMDSC) were associated with reduced OS.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, VU University Medical Center, Cancer Center Amsterdam, Amsterdam, The Netherlands.

ABSTRACT

Background: Cancer-related disturbances in myeloid lineage development, marked by high levels of myeloid-derived suppressor cells (MDSC) and impaired dendritic cell (DC) development, are associated with poor clinical outcome due to immune escape and therapy resistance. Redressing this balance may therefore be of clinical benefit. Here we investigated the effects of combined Prostate GVAX/ipilimumab immunotherapy on myeloid subsets in peripheral blood of castration-resistant prostate cancer (CRPC) patients as well as the putative predictive value of baseline and on-treatment myeloid parameters on clinical outcome.

Methods: Patients with CRPC (n = 28) received thirteen intradermal administrations of Prostate GVAX, consisting of two allogeneic GM-CSF-transduced and irradiated prostate cancer cell lines (LN-CaP and PC3) and six infusions of escalating doses of anti-CTLA4/ipilimumab. Frequencies and activation status of peripheral blood DC (PBDC) and MDSC were determined before, during and after treatment by flowcytometric analysis and related to clinical benefit.

Results: Significant treatment-induced activation of conventional and plasmacytoid DC subsets (cDC and pDC) was observed, which in the case of BDCA1/CD1c(+) cDC1 and MDC8(+)/6-sulfoLacNAc(+) inflammatory cDC3 was associated with significantly prolonged overall survival (OS), but also with the development of autoimmune-related adverse events. High pre-treatment levels of CD14(+)HLA-DR(-)monocytic MDSC (mMDSC) were associated with reduced OS. Unsupervised clustering of these myeloid biomarkers revealed particular survival advantage in a group of patients with high treatment-induced PBDC activation and low pretreatment frequencies of suppressive mMDSC in conjunction with our previously identified lymphoid biomarker of high pretreatment CD4(+)CTLA4(+) T cell frequencies.

Conclusions: Our data demonstrate that DC and MDSC subsets are affected by prostate GVAX/ipilimumab therapy and that myeloid profiling may contribute to the identification of patients with possible clinical benefit of Prostate GVAX/ipilimumab treatment.

No MeSH data available.


Related in: MedlinePlus