Jl. Canon

In vivo stimulation and inhibition of granulopoiesis: the effect of an inflammatory reaction on murine diffusion chamber granulopoiesis.

Humoral factors influencing granulopoiesis have been evaluated using diffusion chambers (DC) implanted in the peritoneal cavity of mice challenged by an aseptic abcess produced by the subcutaneous implantation of copper rods. This resulted in an increase in peripheral blood neutrophils and an increase in tibial granulocytic elements. When DC loaded with bone‐marrow cells were implanted into mice stimulated the day before by an aseptic abcess significantly more CFU‐s, CFU‐c, proliferative and non‐proliferative granulocytes were produced, as compared to DC implanted into control hosts. When DC were implanted 4‐6 d after the induction of inflammation in mice a significant depression of DC granulopoiesis was observed. Levels of serum and DC fluid CSF and serum inhibitors of in vitro colony growth showed no correlation with DC myelopoiesis. The data show that mice undergoing an inflammatory reaction elaborate first humoral substance(s) enhancing CFU‐s and granulocytic growth in DC and next inhibitory factor(s) of DC granulopoiesis.

Letrozole and palbociclib versus chemotherapy as neoadjuvant therapy of high-risk luminal breast cancer

Background Palbociclib is a CDK4/6 inhibitor with demonstrated efficacy and safety in combination with endocrine therapy in advanced luminal breast cancer (LBC). We evaluated the respective efficacy and safety of chemotherapy and letrozole-palbociclib (LETPAL) combination as neoadjuvant treatment in patients with high-risk LBC. Patients and methods NeoPAL (UCBG10/4, NCT02400567) is a randomised, parallel, non-comparative phase II study. Patients with ER-positive, HER2-negative, Prosigna®-defined luminal B, or luminal A and node-positive, stage II-III breast cancer, not candidate for breast-conserving surgery, were randomly assigned to either letrozole (2.5 mg daily) and palbociclib (125 mg daily, 3 weeks/4) during 19 weeks, or to FEC100 (5FU 500 mg/m2, epirubicin 100 mg/m2, cyclophosphamide 500 mg/m2)×3 21-day courses followed by docetaxel 100 mg/m2×3 21-day courses. Primary end point was residual cancer burden (RCB 0-I rate). Secondary end points included clinical response, proliferation-based markers, and safety. Results Overall, 106 patients were randomised [median Prosigna® ROR Score 71 (22-93)]. RCB 0-I was observed in four and eight patients in LETPAL [7.7% (95% CI 0.4-14.9)] and chemotherapy [15.7% (95% CI 5.7-25.7)] arms, respectively. Pathological complete response rates were 3.8% and 5.9%. Clinical response (75%) and breast-conserving surgery rates (69%) were similar in both arms. Preoperative Endocrine Prognostic Index 0 scores (breast cancer-specific survival) were observed in 17.6% and 8.0% of patients in LETPAL and chemotherapy arms, respectively. Safety profile was as expected, with 2 versus 17 serious adverse events (including 11 grade 4 serious AEs in the chemotherapy arm). Conclusion LETPAL combination was associated with poor pathological response but encouraging clinical and biomarker responses in Prosigna®-defined high-risk LBC. Contemporary chemotherapy regimen was associated with poor pathological and biomarker responses, with a much less favourable safety profile. LETPAL combination might represent an alternative to chemotherapy in early high-risk LBC. Clinical Trial Number NCT02400567.

Abstract P1-12-06: UCBG intergroup: 3-years efficacy results of the Unicancer-PACS08 trial including poor prognosis patients treated with docetaxel or ixabepilone in adjuvant setting

Background: Ixabepilone, an epothilone B analog, has demonstrated single-agent activity in metastatic and neoadjuvant settings. The PACS08 trial aimed to compare adjuvant FEC100-Docetaxel regimen to FEC100-Ixabepilone in poor prognosis early breast cancer (BC) composed of patients presenting with triple-negative (TN) [i.e. estrogen receptor (ER)-/progesterone receptor (PR)-/HER2-] or ER+/PR-/HER2- tumor, which are subgroups significantly associated with worse prognosis. Patients and methods: Between 2007 and 2010, 762 patients with unilateral TNBC (n=592, 78%) or node-positive ER+/PR-/HER2- BC (n=170, 22%) were enrolled. Recruitment was interrupted due to BMS decision to stop ixabepilone development in adjuvant setting. Main inclusion criteria were: age Results: Main pts characteristics were well balanced between the 2 arms. As of September 2014, the median follow-up was 36 months. The safety profile indicates that Docetaxel is more often associated to significant haematological toxicities whereas both neurotoxicities and haematological toxicities are reported in Ixabepilone arm. Log-Rank tests indicate no difference between two arms in terms of both DFS and OS (HR=1.2, 95%CI (0.864-1.728), p=0.256 and HR=1.8, 95%CI (0.751-1.855), p=0.473, respectively). Pathological analysis of the PACS08 collection showed that TNBC displayed significantly higher proliferative activity as shown by mitotic count and Ki67 index (p Conclusions: Our results indicate that Ixabepilone doesn9t show higher efficiency compared to Docetaxel in adjuvant setting in poor prognosis early breast cancer. We have an unusual biological collection associated to our clinical data which will allow us to correlate efficacy data to breast cancer subgroups. Other several translational researches are still ongoing. Citation Format: Campone M, Lacroix-Triki M, Roca L, Spielmann M, Wildiers H, Cottu P, Kerbrat P, Levy C, Mayer F, Bachelot T, Wiston T, Eymard J-C, Uwer L, Machiels J-P, Verhoeven D, Jaubert D, Facchini T, Orfeuvre H, Canon J-L, Asselain B, Lemonnier J, Roche H. UCBG intergroup: 3-years efficacy results of the Unicancer-PACS08 trial including poor prognosis patients treated with docetaxel or ixabepilone in adjuvant setting. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-12-06.

Abstract P4-04-08: Presence of tumor-specific cytolytic T cells in human primary breast carcinoma: Consequences for immunotherapy

Background: Immunotherapy through stimulatory antibodies targeting the CTLA-4 or PD-1 pathways has a clear clinical efficacy in a fraction of patients with various cancers. It is likely that the main immune effectors of these therapies are CD8+ cytolytic T lymphocytes (CTL) recognizing tumor-specific antigens. The antigenicity of human tumors has been demonstrated with studies conducted mostly on melanomas. However the genetic mechanisms leading to antigenicity, notably point mutations in the tumor cells, apply to all cancer types. Thus primary breast carcinoma cells do certainly bear tumor-specific antigens, even though the extent of this antigenicity is unknown. Most melanomas, which are highly antigenic tumors, are also immunogenic, i.e. they stimulate spontaneous anti-tumor CTL responses. This immunogenicity, of which the presence of tumor-infiltrating T cells (TILs) is probably a surrogate marker, might be a predictive marker for clinical benefit to immunostimulatory antibodies. Whether primary breast carcinomas are immunogenic is not known, mainly due the absence of autologous tumor cell lines to analyze patients9 T cells. However even in the absence of T-cell aimed immunotherapy the amounts of TILs have been positively correlated with patients9 survival. Here we wished to obtain evidence for the presence of tumor-specific CD8+ T cells in TILs from primary breast carcinomas. Methods: From each tumor we isolated TILs and derived a random set of ±100 CD8+ clones maintained in culture by stimulation with anti-CD3 antibodies, thus irrespective of their antigenic specificity. We screened these clones for recognition of tumor-specific antigens present on the autologous tumor. In the absence of autologous tumor lines we restricted our analysis to mutated antigens selected on the basis of tumor exome sequencing and gene expression profiling. Indels and non-synonymous base substitutions were selected to synthesize candidate mutated peptides. Results: Thus far we have analyzed two hormone receptor-positive HER2-negative primary carcinomas. For one patient we screened 144 T cell clones for recognition of 40 candidate mutated peptides, without any positive result. For the other patient, 6 out of 98 T cell clones recognized 4 out of 119 candidate mutated peptides. Two peptides were recognized by two different T cell clones, i.e. with different T cell receptor sequences. These 4 9antigenic9 mutations appear to be passenger, i.e. the four genes have a low published mutation frequency. Conclusions: We conclude that some human primary breast carcinomas are immunogenic, as one tumor contained at least 6% of tumor-specific T cells among the CD8+ TILs. It suggests that the corresponding patient could benefit from the currently used immunostimulatory antibodies. More work is required to understand the reasons for the negative results in the first patient. We are pursuing the work on 2 HER2-positive and 2 triple-negative tumors, in which TILs are better correlated with prognosis. Our results warrant more investigations on the activation or inhibition of tumor-specific T cells at early stages of human breast cancer development. Citation Format: Schroder DJ, Bricard O, Hames G, Remy N, Carrasco J, Canon J-L, Berliere M, Galant C, Coulie PG. Presence of tumor-specific cytolytic T cells in human primary breast carcinoma: Consequences for immunotherapy. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P4-04-08.

Abstract P3-04-04: Identification of a “BRCAness” signature in triple negative breast cancer by Comparative Genomic Hybridization

Background: Triple Negative Breast Cancers (TNBC) represent 12% to 20% of total Breast Cancers (BC) and have a worse outcome compared with other breast cancer subtypes. TNBC often show a deficiency in DNA double strand break repair mechanisms. This deficiency is generally related to inactivation of a repair enzymatic complex involving BRCA1 caused either by genetic mutations, epigenetic modifications, or by post-transcriptional regulations. The identification of BC presenting a BRCA1 deficiency could be useful to select patients that could benefit from PARP inhibitors, alkylant agents or platinum-based chemotherapy. In this study, we have identified by Comparative Genomic Hybridization array (CGH-array) a recurrent gain in 17q25.3 characteristic of BRCA1 mutated or methylated TNBC. Methods: 130 formalin-fixed paraffin embedded (FFPE) tumours including TNBC (unknown-BRCA1-status, BRCA1 mutated and non-mutated), Luminal A, Luminal B, Her2-neu amplified (Her2+) BC and BRCA1-mutated non-TNBC (mutated-Luminal A, Luminal B, Her2+) were obtained from our local tumour collection. DNA was extracted and genomic copy-number alterations were analysed by CGH-array (Agilent, SuperPrint G3 Human CGH 8×60K Oligo Microarrays). FISH analyses were performed on section from FFPE samples to validate some chromosomal aberrations belonging to the 17q25.3 amplified region evidenced by CGH-array. The study of BRCA1 promoter methylation status in all tumours was carried out by MDxHealth (Liege, Belgium). Results: In this study, we have identified by CGH-array a genomic region (17q25.3) amplified in 90% of the BRCA1 mutated tumours (29/32). This chromosomal gain was studied in other subtypes of BC by CGH-array and it was only evidenced in 30% (6/20) of BRCA1 non-mutated TNBC, 26.67% (4/15) of unknown-BRCA1-status TNBC, 13.64% (3/22) of Luminal B, 19.05% (4/21) of Her2+ and 0% (0/20) of Luminal A breast cancers. FISH assays confirmed these chromosomal amplifications and evidenced like CGH array analyses a significant difference between BRCA1 mutated and non-mutated BC for the 17q25.3 gain. BRCA1 methylation was found only in TNBC (11/58) and was not found in the BRCA1-mutated BC cohort, nor in the Luninal A, Luminal B and Her2+ samples. In BRCA1 non-mutated TNBC, the methylation was found in 8 cases including 4 with 17q25.3 amplification. In the unknown-BRCA1-status TNBC, 3 methylated samples were found with 1 case with co-amplification of the 17q25.3 region. Recurrence of 17q25.3 amplification in BRCA1 mutated tumours as well as its detection in BC having a methylation in BRCA1 promoter suggests that the 17q25.3 gain could be a marker of the BRCA1 deficiency. Identification of relevant genes whose expression is up-regulated within the recurrently gained region is underway. Conclusions: The CGH signature observed in 17q25.3 chromosomal region and FISH assay developed in this study could allow the identification of “BRCAness” breast tumours, improving the diagnostic performance and orienting the selection of the appropriate therapy. The up-regulated genes themselves might also represent potential therapeutic targets. Acknowledgements: This work was financed through the “Plan National Cancer-Action 29” (Belgium) and supported by the UNICANCER-PACS08 trial (France). Citation Information: Cancer Res 2012;72(24 Suppl):Abstract nr P3-04-04.

P5-18-04: Safety Profile of Ixabepilone as Adjuvant Treatment for Poor Prognosis Early Breast Cancer: First Results of the Unicancer-PACS 08 Trial.

Purpose: PACS 01 trial demonstrated that the sequential adjuvant chemotherapy with FEC100 followed by docetaxel (D) significantly improves disease-free and overall survival in node-positive(N+) early breast cancer (BC). However, Triple negative (TN) and ER+/ PR-/HER2− subgroups are significantly associated to a worse prognosis even after adjunction of D. As Ixabepilone (Ixa) has notable preclinical and clinical activity in these subgroups, the PACS 08 trial aims to compare standard FEC100-D regimen to 3 cycles of FEC100 followed by 3 cycles of Ixa. We report the preliminary results of the toxicity profile. Patients and methods: Patients (pts) had localized resectable unilateral ER-/PR-/HER2− or ER+/PR-/HER2− BC. Main inclusion criteria were: age Conclusion: Our results indicate that D arm is more often associated to significant haematological toxicities, whereas both neurotoxicities and haematological toxicities are reported in the Ixa arm. Although significantly more pts discontinued treatment due to adverse events in Ixa arm compared to D arm, Ixa may still represent a promising therapeutic option for pts in the adjuvant setting especially for poor prognosis BC. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-18-04.