Exosomes are endosome-derived nanovesicles actively released into the extracellular environment and biological fluids, both under physiological and pathological conditions, by different cell types. We characterized exosomes constitutively secreted by HER2-overexpressing breast carcinoma cell lines and analyzed in vitro and in vivo their potential role in interfering with the therapeutic activity of the humanized antibody Trastuzumab and the dual tyrosine kinase inhibitor (TKI) Lapatinib anti-HER2 biodrugs. We show that exosomes released by the HER2-overexpressing tumor cell lines SKBR3 and BT474 express a full-length HER2 molecule that is also activated, although to a lesser extent than in the originating cells. Release of these exosomes was significantly modulated by the growth factors EGF and heregulin, two of the known HER2 receptor-activating ligands and naturally present in the surrounding tumor microenvironment. Exosomes secreted either in HER2-positive tumor cell-conditioned supernatants or in breast cancer patients' serum bound to Trastuzumab. Functional assays revealed that both xenogeneic and autologous HER2-positive nanovesicles, but not HER2-negative ones, inhibited Trastuzumab activity on SKBR3 cell proliferation. By contrast, Lapatinib activity on SKBR3 cell proliferation was unaffected by the presence of autologous exosomes. Together, these findings point to the role of HER2-positive exosomes in modulating sensitivity to Trastuzumab, and, consequently, to HER2-driven tumor aggressiveness.

The results are compatible with the hypothesis that the increase in breast cancer risk with increasing BMI among postmenopausal women is largely the result of the associated increase in estrogens, particularly bioavailable estradiol.

Several studies have shown that hormonal, metabolic and inflammatory mechanisms may affect breast cancer progression. We tested the prognostic value of metabolic syndrome in 110 postmenopausal breast cancer patients, who participated in a 1-year dietary intervention study. The risk of adverse events after 5.5 years of follow-up was examined by Cox' proportional hazard modelling, adjusting for hormone receptor status, stage at diagnosis and serum testosterone level, which were shown to significantly affect prognosis. The adjusted hazard ratio of recurrence for the presence of metabolic syndrome at baseline was 3.0 (95% CI 1.2-7.1). Combining metabolic syndrome and serum testosterone, the adjusted hazard ratio of recurrence among women with metabolic syndrome and testosterone levels higher than 0.40 ng/ml (median value) was 6.7 (95% CI 2.3-19.8) compared with that among women without metabolic syndrome and testosterone levels 0.40 ng/ml. The results suggest that metabolic syndrome may be an important prognostic factor for breast cancer. ' 2006 Wiley-Liss, Inc.Key words: breast cancer; recurrences; metabolic syndromeThe prevalence of metabolic syndrome is increasing in parallel with increasing breast cancer incidence worldwide. 1,2 Several studies have suggested that low HDL-cholesterol, 3 high blood glucose, 4 high triglycerides 5 and other aspects of the metabolic syndrome, such as postmenopausal overweight, 1 abdominal obesity, 6,7 hypertension, 8 high levels of insulin and insulin-like growth factor I (IGF-I), 7,9-11 are associated with breast cancer risk. Metabolic and hormonal parameters related to metabolic syndrome have been suggested to affect breast cancer prognosis too. [12][13][14][15] This is the first report addressing the issue whether breast cancer prognosis is affected by metabolic syndrome, defined by 3 or more of the following indicators: fasting glycaemia 110 mg/dl, HDL-cholesterol <50 mg/dl, triglycerides 150 mg/dl, waist circumference 88 cm, systolic pressure 130 mmHg and diastolic pressure 85 mmHg. 16 If confirmed, these results showing a significantly worse prognosis of patients with metabolic syndrome could have important implications for lifestyle intervention to prevent or decelerate cancer progression. Patients and methodsOne hundred and ten postmenopausal women (mean age: 56.8 6 5.6 years) operated for breast cancer since at least a year (4.6 6 4.4 years on average), not undergoing chemotherapy, and with no clinical evidence of disease recurrence, volunteered to participate in a dietary intervention study in which they were requested to follow kitchen courses and modify their diet for 1 year with the aim of reducing insulin and sex hormone levels (The Diana-2 Study). 12 All patients signed an informed consent and the study was approved by the Institutional Review Board and the Ethical Committee of the Milan National Cancer Institute. Results on hormonal changes and the prognostic values of baseline hormone levels (testosterone, oestradiol and insulin) have been published elsewhere. 12,...

Objectives Evidences from either small series or spontaneous reporting are accumulating that SARS-CoV-2 involves the Nervous Systems. The aim of this study is to provide an extensive overview on the major neurological complications in a large cohort of COVID-19 patients. Methods Retrospective, observational analysis on all COVID-19 patients admitted from February 23rd to April 30th, 2020 to ASST Papa Giovanni XXIII, Bergamo, Italy for whom a neurological consultation/neurophysiological assessment/neuroradiologic investigation was requested. Each identified neurologic complication was then classified into main neurologic categories. Results Of 1760 COVID-19 patients, 137 presented neurologic manifestations that manifested after COVID-19 symptoms in 98 pts and was the presenting symptom in 39. Neurological manifestations were classified as: (a) cerebrovascular disease [53 pts (38.7%)] including 37 ischemic and 11 haemorrhagic strokes, 4 transient ischemic attacks, 1 cerebral venous thrombosis; (b) peripheral nervous system diseases [31 (22.6%)] including 17 Guillain-Barrè syndromes; (c) altered mental status [49 (35.8%)] including one necrotizing encephalitis and 2 cases with RT-PCR detection of SARS-Cov-2 RNA in CSF; (d) miscellaneous disorders, among whom 2 patients with myelopathy associated with Ab anti-SARS-CoV-2 in CSF. Patients with peripheral nervous system involvement had more frequently severe ARDS compared to patients with cerebrovascular disease (87.1% vs 42%; difference = 45.1% 95% CI 42.0-48.2; χ 2 = 14.306; p < 0.0002) and with altered mental status (87.1% vs 55.6%; difference = 31.5% 95% CI 27.5-37.5%; χ 2 = 7.055; p < 0.01). Conclusion This study confirms that involvement of nervous system is common in SARS-CoV-2 infection and offers clinicians useful information for prevention and prompt identification in order to set the adequate therapeutic strategies.

Metabolic syndrome (MS), conventionally defined by the presence of at least three out of five dysmetabolic traits (abdominal obesity, hypertension, low plasma HDL-cholesterol, high plasma glucose and high triglycerides), has been associated with an increased risk of several age-related chronic diseases, including breast cancer (BC). This may have prognostic implications for BC survivors. 2,092 early stage BC survivors aged 35-70, recruited in eleven Italian centres 0-5 years after surgical treatment (1.74 years on average), were followed-up over 2.8 years on average for additional BC-related events, including BC-specific mortality, distant metastasis, local recurrences and contralateral BC. At recruitment, 20 % of the patients had MS. Logistic regression models were carried out to generate OR and 95 % confidence intervals (CI) for new BC events associated with MS, adjusting for baseline pathological prognostic factors. New BC events occurred in 164 patients, including 89 distant metastases. The adjusted ORs for women with MS versus women without any MS traits were 2.17 (CI 1.31-3.60) overall, and 2.45 (CI 1.24-4.82) for distant metastasis. The OR of new BC events for women with only one or two MS traits was 1.40 (CI 0.91-2.16). All MS traits were positively associated with new BC events, and significantly so for low HDL and high triglycerides. MS is an important prognostic factor in BC. As MS is reversible through lifestyle changes, interventions to decrease MS traits in BC patients should be implemented in BC clinics.

Because of large intra-individual variation in hormone levels, few studies have investigated the relation of serum sex hormones to breast cancer (BC) in premenopausal women. We prospectively studied this relation, adjusting for timing of blood sampling within menstrual cycle. Premenopausal women (5,963), recruited to the Hormones and Diet in the Etiology of Breast Tumors (ORDET) cohort study, provided a blood sample in the 20 -24th day of their menstrual cycle. The hypothesis that breast cancer (BC) is related to ovarian function dates back over a century. 1 Epidemiological, in vitro and in vivo studies conducted in the second half of the last century made it clear that steroid sex hormones regulate cell proliferation and play a major role in promoting BC. 2,3 Several mechanistic hypotheses for the development of BC have been proposed, 2,4 but until recently, hormone measurements by epidemiological studies have failed to corroborate any of them. Over the last decade, however, several prospective cohort studies in postmenopausal women have shown that BC development is preceded by alterations in levels of circulating sex hormones. 5 High serum levels of free and total estradiol, total testosterone and other estrogens and androgens, as well as low serum levels of sex hormone-binding globulin (SHBG), have been found to be implicated in the risk of BC. 5 Our own study also indicated that high serum levels of free testosterone are associated with the risk of BC. 6 These prospective investigations were carried out with the help of thousands of healthy women who provided blood samples for storage and future nested-in-the-cohort case-control analyses. Compared to case control studies in clinical settings, the strengths of prospective studies are that control subjects belong to the same cohort that generates the incident disease cases and that blood is collected before the diagnosis of cancer thereby excluding abnormal values that may be due to overt illness.Hormone measurements in premenopausal women are difficult to interpret because serum levels change with the menstrual cycle and because cycle length varies inter-and intra-individually. Only a few prospective investigations have addressed the role of sex hormone levels in BC before the menopause; 7-10 all considered small numbers of case women and did not produce clear results. The endocrine basis of BC in premenopause is therefore the subject of several disparate hypotheses. These include the hypothesis of Grattarola, advanced in the 1960s, 11-12 that hyperandrogenism with luteal inadequacy plays a role in the induction of BC, and of Henderson et al. 13 and Key et al. 2 20 years later, that excess of estrogen plus progesterone can be responsible.The present prospective study was designed to investigate whether luteal inadequacy and hyperandrogenism increase the risk of BC in premenopausal women. We collected blood samples from premenopausal women participating in the study on Hormones and Diet in the Etiology of Breast Tumors (ORDET). 6,14 Samples were taken between ...

Results from this prospective study provide evidence for a statistically significant inverse association between melatonin levels, as measured in overnight morning urine, and invasive breast cancer risk in postmenopausal women.

Prospective studies show that high serum levels of androgens and estrogens are associated with increased incidence of postmenopausal breast cancer. The aim of the present analysis was to study the prognostic value of serum testosterone, estradiol and related factors in postmenopausal breast cancer patients. One hundred and ten patients without clinical recurrence were included in the study. After 5.5 years of follow-up, 31 patients developed distant metastasis (16), local relapse (4), or contralateral breast cancer (11). The risk of adverse events in relation to hormone level was examined by Cox' proportional hazard modeling, adjusting for hormone receptor status and stage at diagnosis. Body mass index and serum levels of testosterone, estradiol and glucose were significantly higher in patients who recurred than those who did not. The hazard ratios were 1.8 (95% CI ‫؍‬ 0.5-6.3) for the middle and 7.2 (95% CI ‫؍‬ 2.4 -21.4) for the upper tertiles of baseline testosterone distribution. Other hormones had only minor influence on prognosis. High testosterone predicts breast cancer recurrence. Further studies are required to determine whether dietary or other medical intervention to reduce testosterone can reduce the recurrence of breast cancer.Key words: breast cancer; recurrences; testosterone Prospective cohort studies on healthy volunteers who donated a blood sample at recruitment have shown beyond reasonable doubt that, after menopause, women with high serum levels of steroid sex hormones (both androgens and estrogens) are at increased risk of subsequent breast cancer. Testosterone and estradiol have similar predictive values, with relative risks of the order of 2-3 for women in the highest quintile compared to those in the lowest quintile of the hormone concentration. 1 Several anthropometric and metabolic determinants of high sex hormone availability have also been found associated with breast cancer risk, including obesity, especially abdominal obesity, 2 low serum levels of sex hormone-binding globulin (SHBG), 1 high levels of insulin, 3 fasting glucose and bioavailable insulin-like growth factor-I (IGF-I). 4 Oophorectomy and antiestrogenic treatment reduce breast cancer incidence 5,6 and the incidence of recurrence in breast cancer patients. 7,8 Several studies have also suggested that overweight, 2,9 weight gain during adjuvant treatment, 10 -12 hyperinsulinemia 13 and increased androgenic activity 14 -16 are associated with increased breast cancer recurrences. In the present study, we followed up women enrolled in a previous dietary intervention trial aimed at reducing sex hormone levels in postmenopausal breast cancer patients 17 to examine the relationship between serum hormone levels and cancer recurrence. Material and methodsOne hundred and fifteen postmenopausal women who were operated for breast cancer at least a year previously, not undergoing chemotherapy and with no clinical evidence of disease recurrence volunteered to participate in Diet and Androgens Trial-2 (DIANA-2), a dietary intervention ...