Establishment and Morphological Characterization of Patient-Derived Organoids from Breast Cancer.

BACKGROUND: Patient-derived organoids (PDO) technology represents an emerging tool for the study of tumor biology and drug responsiveness, thus being useful to design personalized medicine approaches. Despite several studies and clinical trials are ongoing using PDO from colorectal and pancreatic cancer, only few research papers have been published exploiting PDO from breast cancer. Here, we have developed a new protocol to establish PDO from surgical and biopsy samples. Furthermore, we have set up also the methodologies adopted for culture and morphological evaluations. RESULTS: Surgical and core biopsy specimens collected from 33 patients with diagnosis of breast cancer have been processed using the protocols here described obtaining PDO from cancerous and healthy mammary tissue (when available) in a quick and easy way with good yields. The more critical aspects influencing the yield were the characteristic of the tissue of origin (healthy vs tumor tissue) and the amount of material obtained after enzymatic digestion process. Success rate from healthy samples was about 20,83%, while this percentage was higher in samples from cancer tissue (i.e. 87,5%). Also the morphological characterization of breast cancer PDO by brightfield and transmission electron microscopy has been reported. CONCLUSIONS: Despite obtaining some organoids from a surgical or biopsy specimen is not a difficult procedure, the establishment of a stable organoid line able to grow and replicate, suitable for long-term biobank storage, is not so obvious. A novel, simple and quick procedure to obtain PDO from surgical and biopsy samples is here proposed to achieve high success rate .

A three-gene signature marks the time to locoregional recurrence in luminal-like breast cancer.

BACKGROUND: Gene expression profiling (GEP)-based prognostic signatures are being rapidly integrated into clinical decision making for systemic management of breast cancer patients. However, GEP remains relatively underdeveloped for locoregional risk assessment. Yet, locoregional recurrence (LRR), especially early after surgery, is associated with poor survival. PATIENTS AND METHODS: GEP was carried out on two independent luminal-like breast cancer cohorts of patients developing early (≤5 years after surgery) or late (>5 years) LRR and used, by a training and testing approach, to build a gene signature able to intercept women at risk of developing early LRR. The GEP data of two in silico datasets and of a third independent cohort were used to explore its prognostic value. RESULTS: Analysis of the first two cohorts led to the identification of three genes, CSTB, CCDC91 and ITGB1, whose expression, derived by principal component analysis, generated a three-gene signature significantly associated with early LRR in both cohorts (P value <0.001 and 0.005, respectively), overcoming the discriminatory capability of age, hormone receptor status and therapy. Remarkably, the integration of the signature with these clinical variables led to an area under the curve of 0.878 [95% confidence interval (CI) 0.810-0.945]. In in silico datasets we found that the three-gene signature retained its association, showing higher values in the early relapsed patients. Moreover, in the third additional cohort, the signature significantly associated with relapse-free survival (hazard ratio 1.56, 95% CI 1.04-2.35). CONCLUSIONS: Our three-gene signature represents a new exploitable tool to aid treatment choice in patients with luminal-like breast cancer at risk of developing early recurrence.

Synovial chondromatosis of the temporomandibular joint: a case description with systematic literature review.

Synovial chondromatosis (SC) of the temporomandibular joint (TMJ) is a rare disease characterized by the presence of calcified loose bodies within the joint, and few systematically gathered data are available about its epidemiology. The aim of this paper was to describe a case of SC of the TMJ, and to carry out a systematic review of the literature on epidemiology over the past decade. A case of a 53-year-old female with the classical triad of signs and symptoms of SC (pain, swelling, restricted mouth opening) is described. A systematic search in the National Library of Medicine's PubMed Database was performed. 155 cases were described in 103 publications. Most dealt with single case reports. Females are affected more than males with a 2.5:1 ratio and the mean age of patients was about 46 years. Late diagnosis is common and in most cases more than 2 years elapsed between symptom onset and surgical intervention. Open TMJ surgery is the treatment of choice, since less invasive techniques, such as arthroscopy, allowed complete removal of the loose bodies only in about half of cases. A single recurrence was described, confirming the benign nature of the disease.

[Microspectrophotometric determination of human leucocyte DNA content in vitro after irradiation]. / Misurazioni microspettrofotometriche dell'ADN di leucociti umani in coltura "in vitro" dopo irradiazione.

Microspectrophotometry was used to determine the DNA content of leucocytes cultured in vitro after irradiation. DNA content values ranges are interpreted as showing the presence of cells with a high ploidy and an increase in the number of cells in G2 stage. These findings are seen as an expression of the arrest of mitosis, without interfering with the duplicative capacity of DNA.

[Microspectrophometric evaluation of DNA as a means of analysis of human leukocyte in vitro proliferative kinetics]. / Analisi della cinetica proliferativa dei leucociti umani in coltura "in vitro" per mezzo della valutazione microspettrofotometrica dell'ADN.

Microspectrophotometric determination of in vitro human leucocyte DNA content, with or without PHA stimulation, is reported. The results obtained are compared with those observed with tritiated thymidine and their parallel pattern makes in clear that DNA microspectrophotometry can be employed in the study of proliferative kinetics.

Clastogenic effect of BrdU on different phases of human lymphocyte "in vitro" cell cycle.

From the data presented, it can be pointed out a mitotic delay in the cell cycle, possibly due to BrdU cytotoxic effect. Irradiation in G1 phase does not produce any increase in SCE frequency while irradiation in S or G2 phase increases significantly this frequency. Our data support the hypothesis that BrdU does not produce by itself an increase of chromosome aberrations but act as sensitizer for cells which have incorporated it.

Analysis of cell kinetics, chromosome aberration frequency and sister chromatid exchanges in lymphocyte "in vitro" cultures of patients irradiated by 131I. Preliminary note.

In the present note we analyse the possible effect of 131I administration on lymphocyte cultures or patients previously subjected to thyroidectomy because of carcinoma. Three parameters were taken into consideration: cell kinetics, SCE frequency and chromosome aberrations frequency before and after treatment. It is possible to point out only an increase of chromosome aberrations. Our data stress the importance of considering several variables at the same time when we are faced with the problem of evaluating possible DNA damage.

Analysis of chromosome aberrations after exposure to ionizing radiation in "in vitro" cultures of human lymphocytes by means of BrdU incorporation.

From the data presented, it can be pointed out that a linear relationship does not exist between irradiation doses and Sister chromatid exchanges. The number of chromosome aberrations instead increases regularly in correspondence with the increase of the irradiation doses while SCE remain at the same level. Our data demonstrate that, at each division, there is, at least in vitro, the loss of about 50% of the dicentrics and support the necessity in order to evaluate chromosome damage, of considering cultures harvested not later than 44 hours.