BackgroundTriple-negative breast cancer (TNBC) treatment is currently restricted to chemotherapy. Hence, tumor-specific molecular targets and/or alternative therapeutic strategies for TNBC are urgently needed. Immunotherapy is emerging as an exciting treatment option for TNBC patients. The aspartic protease cathepsin D (cath-D), a marker of poor prognosis in breast cancer (BC), is overproduced and hypersecreted by human BC cells. This study explores whether cath-D is a tumor cell-associated extracellular biomarker and a potent target for antibody-based therapy in TNBC.MethodsCath-D prognostic value and localization was evaluated by transcriptomics, proteomics and immunohistochemistry in TNBC. First-in-class anti-cath-D human scFv fragments binding to both human and mouse cath-D were generated using phage display and cloned in the human IgG1 λ format (F1 and E2). Anti-cath-D antibody biodistribution, antitumor efficacy and in vivo underlying mechanisms were investigated in TNBC MDA-MB-231 tumor xenografts in nude mice. Antitumor effect was further assessed in TNBC patient-derived xenografts (PDXs).ResultsHigh CTSD mRNA levels correlated with shorter recurrence-free survival in TNBC, and extracellular cath-D was detected in the tumor microenvironment, but not in matched normal breast stroma. Anti-cath-D F1 and E2 antibodies accumulated in TNBC MDA-MB-231 tumor xenografts, inhibited tumor growth and improved mice survival without apparent toxicity. The Fc function of F1, the best antibody candidate, was essential for maximal tumor inhibition in the MDA-MB-231 model. Mechanistically, F1 antitumor response was triggered through natural killer cell activation via IL-15 upregulation, associated with granzyme B and perforin production, and the release of antitumor IFNγ cytokine. The F1 antibody also prevented the tumor recruitment of immunosuppressive tumor-associated macrophages M2 and myeloid-derived suppressor cells, a specific effect associated with a less immunosuppressive tumor microenvironment highlighted by TGFβ decrease. Finally, the antibody F1 inhibited tumor growth of two TNBC PDXs, isolated from patients resistant or not to neo-adjuvant chemotherapy.ConclusionCath-D is a tumor-specific extracellular target in TNBC suitable for antibody-based therapy. Immunomodulatory antibody-based strategy against cath-D is a promising immunotherapy to treat patients with TNBC.Electronic supplementary materialThe online version of this article (10.1186/s40425-019-0498-z) contains supplementary material, which is available to authorized users.

a b s t r a c tThe analysis of phytoliths, pollen, charcoal and other macroremains was carried out in the neolithic shelter of ''La Grande Rivoire'', Vercors massif (French Alps). The results show the predominance of tree species, in the form of phytoliths, clustered pollen, stomata, small branches charcoal, needles, bark, buds. The practice of leaf fodder is already known in the alpine and circum-alpine area from archaeological and historical sources. The analyses of the neolithic dung levels of ''La Grande Rivoire'' illustrate the use of leafy and flowering tree branches as fodder. The results also suggest that some species were used for special purpose in relation with the tending of livestock (litter, dietary supplement, veterinary practices).

This paper aims to define the first chrono-cultural framework on the domestication and early diffusion of the opium poppy using small-sized botanical remains from archaeological sites, opening the way to directly date minute short-lived botanical samples. We produced the initial set of radiocarbon dates directly from the opium poppy remains of eleven Neolithic sites (5900–3500 cal BCE) in the central and western Mediterranean, northwestern temperate Europe, and the western Alps. When possible, we also dated the macrobotanical remains originating from the same sediment sample. In total, 22 samples were taken into account, including 12 dates directly obtained from opium poppy remains. The radiocarbon chronology ranges from 5622 to 4050 cal BCE. The results show that opium poppy is present from at least the middle of the sixth millennium in the Mediterranean, where it possibly grew naturally and was cultivated by pioneer Neolithic communities. Its dispersal outside of its native area was early, being found west of the Rhine in 5300–5200 cal BCE. It was introduced to the western Alps around 5000–4800 cal BCE, becoming widespread from the second half of the fifth millennium. This research evidences different rhythms in the introduction of opium poppy in western Europe.