Efficacy and safety of avapritinib in previously treated patients with advanced systemic mastocytosis.

Advanced systemic mastocytosis (AdvSM) is a rare myeloid neoplasm, driven by the KIT D816V mutation in >90% of patients. Avapritinib, a potent, highly selective D816V-mutant KIT inhibitor, is approved for treatment of adults with AdvSM by the US Food and Drug Administration, regardless of prior therapy, and the European Medicines Agency for patients with prior systemic therapy, based on EXPLORER (#NCT02561988; clinicaltrials.gov) and PATHFINDER (#NCT03580655; clinicaltrials.gov) clinical studies. We present latest pooled efficacy and safety analyses from patients who received ≥1 systemic therapy prior to avapritinib in EXPLORER/PATHFINDER. Overall response rate in response-evaluable patients (n = 31) was 71% (95% confidence interval: 52% to 86%; 22/31), including 19% (6/31) with complete remission (CR)/CR with partial recovery of peripheral blood counts (CRh). Median time to response was 2.3 months, median time to CR/CRh was 7.4 months, and median duration of response (DOR) was not reached. Reductions ≥50% in bone marrow mast cell infiltration (89%), KIT D816V variant allele fraction (66%), serum tryptase (89%), and reductions ≥35% in spleen size (70%) occurred in most patients. Median OS was not reached (median follow-up 17.7 months). Avapritinib was effective in all AdvSM subtypes, regardless of number/type of prior therapies or poor prognostic somatic mutations. Treatment-related adverse events (TRAEs) were observed in 94% of patients, most commonly grade 1/2; 57% had TRAEs of at least grade 3; 81% remained on treatment at 6 months. Avapritinib in adults with AdvSM who received prior systemic therapy was generally well tolerated, with high response rates regardless of prior systemic therapy.

Mineralogical Controls on the Bioaccessibility of Arsenic in Fe(III)-As(V) Coprecipitates.

X-ray amorphous Fe(III)-As(V) coprecipitates are common initial products of oxidative As- and Fe-bearing sulfide weathering, and often control As solubility in mine wastes or mining-impacted soils. The formation conditions of these solids may exert a major control on their mineralogical composition and, hence, As release in the gastric tract of humans after incidental ingestion of As-contaminated soil. Here, we synthesized a set of 35 Fe(III)-As(V) coprecipitates as a function of pH (1.5-8) and initial molar Fe/As ratio (0.8-8.0). The solids were characterized by synchrotron X-ray diffraction, FT-IR spectroscopy, and electrophoretic mobility measurements, and their As bioaccessibility (BAAs) was evaluated using the gastric-phase Solubility/Bioavailability Research Consortium in vitro assay (SBRC-G). The coprecipitates contained 1.01-4.51 mol kg-1 As (molar Fe/Assolid: 1.00-8.29) and comprised varying proportions of X-ray amorphous hydrous ferric arsenates (HFAam) and As(V)-adsorbed ferrihydrite. HFAam was detected up to pH 6 and its fraction decreased with increasing pH and molar Fe/As ratio. Bioaccessible As ranged from 2.9 to 7.3% of total As (x̅ = 4.8%). The BAAs of coprecipitates formed at pH ≤ 4 was highest at formation pH 3 and 4 and controlled by the intrinsically high solubility of the HFAam component, possibly enhanced by sorbed sulfate. In contrast, the BAAs of coprecipitates dominated by As(V)-adsorbed ferrihydrite was much lower and controlled by As readsorption and/or surface precipitation in the gastric fluid. Bioaccessible As increased up to 95% with increasing liquid-to-solid ratio, indicating an enhanced solubility of these solids due to interactions between Fe and the glycine buffer. We conclude (i) that natural Fe(III)-As(V) coprecipitates exhibit a particularly high solubility in the human gastric tract when formed at pH ∼ 3-4 in the presence of sulfate, and (ii) that the in vitro bioaccessibility of As in Fe(III)-As(V) coprecipitates as assessed by tbe SBRC-G assay depends critically on their solid-phase concentration in As-contaminated soil and mine-waste materials.

Effects of Manganese Oxide on Arsenic Reduction and Leaching from Contaminated Floodplain Soil.

Reductive release of the potentially toxic metalloid As from Fe(III) (oxyhydr)oxides has been identified as an important process leading to elevated As porewater concentrations in soils and sediments. Despite the ubiquitous presence of Mn oxides in soils and their oxidizing power toward As(III), their impact on interrelated As, Fe, and Mn speciation under microbially reducing conditions remains largely unknown. For this reason, we employed a column setup and X-ray absorption spectroscopy to investigate the influence of increasing birnessite concentrations (molar soil Fe-to-Mn ratios: 4.8, 10.2, and 24.7) on As speciation and release from an As-contaminated floodplain soil (214 mg As/kg) under anoxic conditions. Our results show that birnessite additions significantly decreased As leaching. The reduction of both As and Fe was delayed, and As(III) accumulated in birnessite-rich column parts, indicating the passivation of birnessite and its transformation products toward As(III) oxidation and the precipitation of Fe(III)(oxyhydr)oxides. Microbial Mn reduction resulted in elevated soil pH values, which in turn lowered the microbial activity in the birnessite-enriched soil. We conclude that in Mn-oxide-rich soil environments undergoing redox fluctuations, the enhanced As adsorption to newly formed Fe(III) (oxyhydr)oxides under reducing conditions leads to a transient stabilization of As.

Impact of birnessite on arsenic and iron speciation during microbial reduction of arsenic-bearing ferrihydrite.

Elevated solution concentrations of As in anoxic natural systems are usually accompanied by microbially mediated As(V), Mn(III/IV), and Fe(III) reduction. The microbially mediated reductive dissolution of Fe(III)-(oxyhydr)oxides mainly liberates sorbed As(V) which is subsequently reduced to As(III). Manganese oxides have been shown to rapidly oxidize As(III) and Fe(II) under oxic conditions, but their net effect on the microbially mediated reductive release of As and Fe is still poorly understood. Here, we investigated the microbial reduction of As(V)-bearing ferrihydrite (molar As/Fe: 0.05; Fe tot: 32.1 mM) by Shewanella sp. ANA-3 (10(8) cells/mL) in the presence of different concentrations of birnessite (Mn tot: 0, 0.9, 3.1 mM) at circumneutral pH over 397 h using wet-chemical analyses and X-ray absorption spectroscopy. Additional abiotic experiments were performed to explore the reactivity of birnessite toward As(III) and Fe(II) in the presence of Mn(II), Fe(II), ferrihydrite, or deactivated bacterial cells. Compared to the birnessite-free control, the highest birnessite concentration resulted in 78% less Fe and 47% less As reduction at the end of the biotic experiment. The abiotic oxidation of As(III) by birnessite (k initial = 0.68 ± 0.31/h) was inhibited by Mn(II) and ferrihydrite, and lowered by Fe(II) and bacterial cell material. In contrast, the oxidation of Fe(II) by birnessite proceeded equally fast under all conditions (k initial = 493 ± 2/h) and was significantly faster than the oxidation of As(III). We conclude that in the presence of birnessite, microbially produced Fe(II) is rapidly reoxidized and precipitates as As-sequestering ferrihydrite. Our findings imply that the ability of Mn-oxides to oxidize As(III) in water-logged soils and sediments is limited by the formation of ferrihydrite and surface passivation processes.

Treatment of advanced metastasized breast cancer with bone marrow-derived tumour-reactive memory T cells: a pilot clinical study.

BACKGROUND: Breast cancer patients frequently harbour tumour-reactive memory T cells in their bone marrow (BM) but not in the blood. After reactivation ex-vivo these cells rejected autologous breast tumours in xenotransplanted mice demonstrating therapeutic potential upon reactivation and mobilization into the blood. We conducted a clinical pilot study on metastasized breast cancer patients to investigate if ex-vivo reactivation of tumour-reactive BM memory T cells and their adoptive transfer is feasible and increases the frequencies of tumour-reactive T cells in the blood. METHODS: The study protocol involved one transfusion of T cells which were reactivated in vitro with autologous dendritic cells pulsed with lysate of MCF7 breast cancer cells as source of tumour antigens. Immunomonitoring included characterization of T cell activation in vitro and of tumour-specific T cells in the blood by interferon (IFN)-gamma ELISPOT assay, HLA-tetramers and antigen-induced interleukin (IL)-4 secretion. RESULTS: Twelve patients with pre-existing tumour-reactive BM memory T cells were included into the study. In all cases, the treatment was feasible and well tolerated. Six patients (responders) showed by ELISPOT assay de-novo tumour antigen-specific, IFN-gamma-secreting T cells in the blood after 7 days. In contrast, non responders showed in the blood tumour antigen-induced IL-4 responses. All responders received more than 6.5 x 10(3) tumour-reactive T cells. In contrast, all non responders received lower numbers of tumour antigen-reactive T cells. This was associated with reduced activation of memory T cells in activation cultures, increased amounts of CD4(+) CD25(high) regulatory T cells in the BM and increased tumour antigen-dependent IL-10 secretion. The latter was prevented by preceding depletion of regulatory T cells suggesting that regulatory T cells in the BM can inhibit reactivation of tumour-specific T cells. CONCLUSION: Taken together, adoptive transfer of ex-vivo re-stimulated tumour-reactive memory T cells from BM of metastasized breast cancer patients can induce the presence of tumour antigen-reactive type-1 T cells in the peripheral blood.

Melanoma-reactive T cells in the bone marrow of melanoma patients: association with disease stage and disease duration.

Using IFN-gamma enzyme-linked immunospot, we investigated reactivity of T cells from bone marrow and peripheral blood to melanoma lysate-pulsed autologous dendritic cells in 40 melanoma patients. Melanoma-reactive T cells were present in the bone marrow of seven patients and in peripheral blood of four patients. In the bone marrow, melanoma-reactive T cells were present in 6 of 21 stage IV patients and in 1 of 10 stage III patients, whereas none were detected in stage I to II patients (0 of 9). The occurrence of tumor-reactive bone marrow T cells in melanoma patients was associated with advanced disease stage, disease duration and tumor load, and independent of treatment. These findings provide new insights into the generation of T-cell responses in melanoma patients.

Specifically activated memory T cell subsets from cancer patients recognize and reject xenotransplanted autologous tumors.

Bone marrow of breast cancer patients was found to contain CD8(+) T cells specific for peptides derived from breast cancer-associated proteins MUC1 and Her-2/neu. Most of these cells had a central or effector memory phenotype (CD45RA(-)CD62L(+) or CD45RA(-)CD62L(-), respectively). To test their in vivo function, we separated bone marrow-derived CD45RA(+) naive or CD45RA(-)CD45RO(+) memory T cells, stimulated them with autologous dendritic cells pulsed with tumor lysate, and transferred them into NOD/SCID mice bearing autologous breast tumors and normal skin transplants. CD45RA(-) memory but not CD45RA(+) naive T cells infiltrated autologous tumor but not skin tissues after the transfer. These tumor-infiltrating cells had a central or effector memory phenotype and produced perforin. Many of them expressed the P-selectin glycoprotein ligand 1 and were found around P-selectin(+) tumor endothelium. Tumor infiltration included cluster formation in tumor tissue by memory T cells with cotransferred dendritic cells. It was associated with the induction of tumor cell apoptosis and significant tumor reduction. We thus demonstrate selective homing of memory T cells to human tumors and suggest that tumor rejection is based on the recognition of tumor-associated antigens on tumor cells and dendritic cells by autologous specifically activated central and effector memory T cells.