In breast cancer (BC) patients, local recurrences often arise in proximity of the surgical scar, suggesting that response to surgery may have a causative role. Radiotherapy (RT) after lumpectomy ...significantly reduces the risk of recurrence. We investigated the direct effects of surgery and of RT delivered intraoperatively (IORT), by collecting irradiated and non-irradiated breast tissues from BC patients, after tumor removal. These breast tissue specimens have been profiled for their microRNA (miR) expression, in search of differentially expressed miR among patients treated or not with IORT. Our results demonstrate that IORT elicits effects that go beyond the direct killing of residual tumor cells. IORT altered the wound response, inducing the expression of miR-223 in the peri-tumoral breast tissue. miR-223 downregulated the local expression of epidermal growth factor (EGF), leading to decreased activation of EGF receptor (EGFR) on target cells and, eventually, dampening a positive EGF-EGFR autocrine/paracrine stimulation loop induced by the post-surgical wound-healing response. Accordingly, both RT-induced miR-223 and peri-operative inhibition of EGFR efficiently prevented BC cell growth and reduced recurrence formation in mouse models of BC. Our study uncovers unknown effects of RT delivered on a wounded tissue and prompts to the use of anti-EGFR treatments, in a peri-operative treatment schedule, aimed to timely treat BC patients and restrain recurrence formation.
Locally advanced HER2-overexpressing breast cancer (BC) patients achieve a high rate of pathological complete responses (pCR) after neoadjuvant chemotherapy (NC). The apparently unaltered immune ...proficiency of these patients together with the immune-modulating activities of NC drugs suggest a potential contribution of host immunity in mediating clinical responses. We thus performed an extensive immunomonitoring in locally advanced BC patients undergoing NC to identify immunological correlates of pCR induction.
The immune profile of 40 HER2-positive and 38 HER2-negative BC patients was characterized at diagnosis and throughout NC (Paclitaxel and Trastuzumab, or Docetaxel and Epirubicin, respectively). The percentages of circulating immune cell subsets including T and B lymphocytes, Natural Killer (NK) cells, regulatory T cells, T helper 17 lymphocytes, were quantified by multiparametric flow cytometry. NK cells functional activity was evaluated through the analysis of NF-kB nuclear translocation by Multispectral flow cytometry, and with the in vitro monitoring of Trastuzumab-mediated antibody-dependent cell cytotoxicity (ADCC). CD8(+) T cell responses against six different tumor-associated antigens (TAA) were characterized by IFN-γ ELISPOT and IFN-γ/IL-2 DualSpot assays.
After NC, HER2-positive patients showed a significant increase in the number of NK cells and regulatory T cells irrespective of the pathological response, whereas patients undergoing a pCR disclosed higher percentages of T helper 17 cells. Notably, a significant increase in the number of activated NK cells was observed only in HER2-positive patients achieving a pCR. Characterization of anti-tumor T cell responses highlighted sustained levels of CD8(+) T cells specific for survivin and mammaglobin-A throughout NC in patients undergoing a pCR in both arms. Moreover, HER2-positive patients achieving a pCR were characterized by a multi-epitopic and polyfunctional anti-tumor T cell response, markedly reduced in case of partial response.
These results indicate that maintenance of functional T cell responses against selected antigens and improvement of NK cell proficiency during NC are probably critical requirements for pCR induction, especially in HER2-positive BC patients. Trail registration:
NCT02307227, registered on ClinicalTrials.gov ( http://www.clinicaltrials.gov , November 26, 2014).
Background: Triple Negative Breast Cancer (TNBC) is an aggressive malignancy with dismal prognosis owing to high levels of recurrence. It has been shown that p7036 kinase (p7056K) is activated via ...post-surgical inflammation and can mediate recurrence in a mouse model of TNBC. The purpose of the current study is to validate whether a novel p70S6K1 inhibitor, FS115, can control the formation of recurrence and metastasis in pre-clinical models of TNBC. Material and Methods: FS115 was synthesized by Sentinel Oncology. IC50s were determined via radiometric kinase assays. Cell colony assay: MDA-MB-231 cells (basal, TNBC) were treated with FS115 or vehicle then seeded onto plates and incubated for 14 d prior to colony counting. The pharmacokinetic (PK) profile was determined in two separate cohorts of CD-i mice (dosed P0 or IV with FSI15). Pharmacodynamics (PD) were determined as follows: nude mice bearing MDA-MB-231 tumours were dosed with FS115 (125mg/kg BID x 3 P0) or vehicle, then 12 h after the last dose mice were sacrificed and tumour ELISA carried out to measure levels of p-S6 240/244, t-S6, p-AKT 473 and t-AKT. Local recurrence was modelled in vivo: MDA-MB-231 orthotopic primary tumors were first grown in nude mice. The mice were treated with FS115 or vehicle in a pen-operative schedule (day -1, day 0, day +1; surgery on day 0) then monitored for recurrence for 56 d. Metastasis was modelled in vivo as follows: On day -1, nude mice were assigned to treatment groups. On day 0, mice were given an intracardiac injection of MDA-MB-231-Iuc cells (luciferase-expressing). Mice were dosed for a further 68 days and were then imaged for total photon flux to show metastatic spread. Results: In vitro, FS1 15 potently inhibits p70S6K1 (IC50 0.035 μM) with high selectivity over P13K pathway kinases (AKT2 IC50 23.8 μM). In MDA- MB-231, a TNBC cell line, FS115 inhibits p70S6K activity in a dose- dependent manner (optimal effect at 10 μM) and suppresses colony growth in the range 1-10μm. The PK profile of FS115 in mice shows high oral bioavailability (>95%) and favorable brain penetration (4:1 B:P). PD studies reveal that FS115 inhibits phosphorylation of p70S6K substrate S6 in tumour by 55% without concomitant AKT activation. In mouse models of TNBC, FS115 was found to inhibit multiple facets of the disease including (a) tumor take rate and growth, (b) local recurrence and (c) metastasis, including brain metastasis. Conclusions: Here, we show that a small molecule oral inhibitor of p70S6K1, FS115, dosed to mice in a pen-operative schedule was effective in decreasing local recurrence of breast cancer and in long-term treatment schedule was well-tolerated and efficiently suppressed distant metastasis formation. Altogether, these findings suggest that an inhibitor of p70S6K1 could provide a targeted treatment option for TNBC patients at high risk of recurrence.
Purpose:
EBT2 radiochromic films were studied and used forin vivo dosimetry in targeted intraoperative radiotherapy (TARGIT), a technique in which the Intrabeam system (Carl Zeiss, Oberkochen, ...Germany) is used to perform intraoperative partial breast irradiation with x-rays of 50 kVp.
Methods:
The energy of the radiation emitted by the Intrabeam with the different spherical applicators, under 1 and 2 cm of solid water, and under the tungsten impregnated rubber used for shielding of the heart in TARGIT of the breast, was characterized with measurements of half-value layer (HVL). The stability of response of EBT2 was verified inside this range of energies. EBT2 films were calibrated using the red and green channels of the absorption spectrum in the 0–20 Gy dose range delivered by the Intrabeam x-rays. The dependence of film response on temperature during irradiation was measured. Forin vivo dosimetry, pieces of radiochromic films wrapped in sterile envelopes were inserted after breast conserving surgery and before TARGIT into the excision cavity, on the skin and on the shielded pectoralis fascia for treatments of the left breast.
Results:
HVLs of the Intrabeam in TARGIT of the breast correspond to effective energies of 20.7–36.3 keV. The response of EBT2 was constant inside this range of energies. We measured the dose to the target tissue and to organs at risk in 23 patients and obtained an average dose of 13.52 ± 1.21 Gy to the target tissue. Dose to the skin in close proximity to the applicator was 2.22 ± 0.97 Gy, 0.29 ± 0.17 Gy at 5–10 cm from the applicator, and 0.08 ± 0.07 Gy at more than 10 cm from the applicator. Dose to the pectoral muscle for left breast treatment was 0.57 ± 0.23 Gy.
Conclusions:
Our results show that EBT2 films are accurate at the beam energies, dose range, and irradiation temperature found in TARGIT and thatin vivo dosimetry in TARGIT with EBT2 films wrapped in sterile envelopes is a feasible procedure. Measured dose to the organs at risk indicates that the technique is safe from side effects to the skin and the heart.
To report cross-sectionally serum levels of 25-hydroxyvitamin D 25(OH)D in women living in Italy within 12 months from breast cancer (BC) diagnosis.
Baseline data were obtained from 394 women ...diagnosed with primary BC, enrolled from 2016 to 2019 in a lifestyle trial conducted in Italy. Subjects' characteristics were compared between two 25(OH)D concentrations (hypovitaminosis D<20 and ≥20 ng/mL) with the Chi-squared test or Fisher's exact test for small-expected counts. Using multiple logistic regression-adjusted models, we estimated odds ratios (ORs) of hypovitaminosis D with 95% confidence intervals (CIs) in the total sample and in the unsupplemented subgroup.
Hypovitaminosis D was found in 39% of all subjects, 60% in unsupplemented subjects, and 10% in supplemented subjects. Increasing ORs of hypovitaminosis D were found with increasing body mass index, 25-30, >30, and ≥35 versus <25 kg/m2 (ORs: 2.50, 4.64, and 5.81, respectively, in the total cohort and ORs: 2.68, 5.38, and 7.08 in the unsupplemented); living in the most southern Italian region (OR 2.50, 95%CI 1.22-5.13); and with hypertriglyceridemia (OR 2.46; 95%CI 1.16-5.22), chemotherapy history (OR 1.86, 95%CI 1.03-3.38), and inversely with anti-estrogenic therapy (OR 0.43, 95%CI 0.24-0.75) in the total sample.
Hypovitaminosis D in women recently diagnosed with BC and participating in a lifestyle trial in Italy was widespread and highest with obesity, hypertriglyceridemia, and chemotherapy use. Considering that hypovitaminosis D is a risk factor for lower efficacy of bone density treatments and possibly BC mortality, our results suggest the need to promptly address and treat vitamin D deficiency.
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