Mitotic cell death induction by targeting the mitotic spindle with tubulin-inhibitory indole derivative molecules

Tubulin-targeting molecules are widely used cancer therapeutic agents. They inhibit microtubule-based structures, including the mitotic spindle, ultimately preventing cell division. The final fates of microtubule-inhibited cells are however often heterogeneous and difficult to predict. While recent work has provided insight into the cell response to inhibitors of microtubule dynamics (taxanes), the cell response to tubulin polymerization inhibitors remains less well characterized. Arylthioindoles (ATIs) are recently developed tubulin inhibitors. We previously identified ATI members that effectively inhibit tubulin polymerization in vitro and cancer cell growth in bulk cell viability assays. Here we characterise in depth the response of cancer cell lines to five selected ATIs. We find that all ATIs arrest mitotic progression, yet subsequently yield distinct cell fate profiles in time-lapse recording assays, indicating that molecules endowed with similar tubulin polymerization inhibitory activity in vitro can in fact display differential efficacy in living cells. Individual ATIs induce cytological phenotypes of increasing severity in terms of damage to the mitotic apparatus. That differentially triggers MCL-1 down-regulation and caspase-3 activation, and underlies the terminal fate of treated cells. Collectively, these results contribute to define the cell response to tubulin inhibitors and pinpoint potentially valuable molecules that can increase the molecular diversity of tubulin-targeting agents

New Indole Tubulin Assembly Inhibitors Cause Stable Arrest of Mitotic Progression, Enhanced Stimulation of Natural Killer Cell Cytotoxic Activity, and Repression of Hedgehog-Dependent Cancer

We designed 39 new 2-phenylindole derivatives as potential anticancer agents bearing the 3,4,5-trimethoxyphenyl moiety with a sulfur, ketone, or methylene bridging group at position 3 of the indole and with halogen or methoxy substituent(s) at positions 4-7. Compounds 33 and 44 strongly inhibited the growth of the P-glycoprotein-overexpressing multi-drug-resistant cell lines NCI/ADR-RES and Messa/Dx5. At 10 nM, 33 and 44 stimulated the cytotoxic activity of NK cells. At 20-50 nM, 33 and 44 arrested >80% of HeLa cells in the G2/M phase of the cell cycle, with stable arrest of mitotic progression. Cell cycle arrest was followed by cell death. Indoles 33, 44, and 81 showed strong inhibition of the SAG-induced Hedgehog signaling activation in NIH3T3 Shh-Light II cells with IC50 values of 19, 72, and 38 nM, respectively. Compounds of this class potently inhibited tubulin polymerization and cancer cell growth, including stimulation of natural killer cell cytotoxic activity and repression of Hedgehog-dependent cancer

HETEROTOPIC ERYTHROPOIESIS IN LIVER TRANSPLANTATION

Various conditions may restore hematopoietic activity in the adult liver. Of them, liver transplantation is of most interest. The causes and the mechanism of hematopoiesis restoral in liver grafts is not yet fully clarified. The Authors analyze the incidence of immature erythroid cells in a series of hepatic fine-needle aspiration biopsies performed in 28 patients with orthotopic liver transplant and in two clusters. The post-mortem liver examination, available in the two clusters, confirmed the ungoing hematopoietic activity

Oral contraceptives and clinical recurrence of human papillomavirus lesions and cervical intraepithelial neoplasia following treatment

Objective: To evaluate the effect of oral contraceptive use on the recurrence rate of human papillomavirus (HPV) lesions and cervical intraepithelial neoplasia (CIN) following ablative or excisional procedures in a long-term follow-up. Method: The study was conducted with 650 oral contraceptive users presenting with HPV lesions and/or CIN, and 670 women who had these lesions but did not use oral contraceptives acted as controls. The participants underwent cytologic evaluations, colposcopy, and direct biopsy, followed by either ablative treatment by laser carbon dioxide vaporization or excision by a loop electrosurgical excision procedure or cold knife conization. They were then followed up for a minimum of 5 years. Results: The recurrence rates did not differ statistically between the case and control groups. Conclusion: Oral contraceptive use was not found to increase the recurrence rate of HPV lesions and/or CIN after ablative or surgical treatment. © 2007 International Federation of Gynecology and Obstetrics

Clinical management and follow-up of squamous intraepithelial cervical lesions during pregnancy and postpartum

Background: The incidence of cervical cancer in pregnancy is estimated to be 1-10/10000 pregnancies. Approximately 3% of cervical cancers are diagnosed during pregnancy. The incidence of abnormal Pap smears has been reported to be 5%-8%. Data on the spontaneous evolution of an intraepithelial neoplasia during pregnancy are quite diverse. Of dysplasia cases diagnosed during pregnancy, 10%-70% regress and sometimes even disappear postpartum, while persistence in the severity of cervical neoplasia is reported in 25%-47% and progression occurs in 3%-30%. However, adequate follow-up and definitive management in the postpartum period is important. The objective of the study was to assess proper management of squamous intraepithelial lesion (SIL) during and after pregnancy, to assess regression, persistence and risk of progression and the predictive role of HPV tests. Materials and Methods: Thirty-one out of 721 pregnant women with a diagnosis of low- and high-grade SIL were observed. All patients were triaged using standard colposcopy. The histological diagnosis was assessed by colposcopic direct biopsies. In patients affected by high-SIL with colposcopic findings of suspected micro-invasive lesions, a loop electrosurgical excisional procedure (LEEP) was carried out in pregnancy. High risk HPV tests were performed using PCR. The patients were followed up with cytology and colposcopy every 6-8 weeks during gestation and nine weeks postpartum. They were re-evaluated using cytology, colposcopy and histology for a final diagnosis and, when necessary, submitted to treatment. The patients were followed up for a minimum of 5 years. The HPV test was performed once at 6-8 weeks during gestation and annually during the follow-up. Results: Of the 31 patients with abnormal cytology, histological analysis revealed 10 cervical intraepithelial neoplasia (CIN) 1, 5 CIN 2 and 16 CIN 3. The HPV test at diagnosis was positive for HPV 16 type in 22 cases and negative in 9. Five patients with CIN 2 and 11 with CIN 3 were followed up; 5 patients with CIN 3 with colposcopic findings of suspected microinvasive lesions were submitted to an excisional procedure with LEEP before the 16th week of pregnancy. Conclusion: Performing high-risk HPV tests may improve the follow-up of patients with SIL in pregnancy and postpartum in addition to cytology and colposcopy to indicate persistence/progression of the lesions. Proper management and adequate follow-up could be proposed in pregnancy and postpartum

Three de novo DDX3X variants associated with distinctive brain developmental abnormalities and brain tumor in intellectually disabled females

De novo DDX3X variants account for 1–3% of syndromic intellectual disability (ID) in females and have been occasionally reported in males. Furthermore, somatic DDX3X variants occur in several aggressive cancers, including medulloblastoma. We report three unrelated females with severe ID, dysmorphic features, and a common brain malformative pattern characterized by malformations of cortical development, callosal dysgenesis, basal ganglia anomalies, and midbrain–hindbrain malformations. A pilocytic astrocytoma was incidentally diagnosed in Patient 1 and trigonocephaly was found in Patient 2. With the use of family based whole exome sequencing (WES), we identified three distinct de novo variants in DDX3X. These findings expand the phenotypic spectrum of DDX3X-related disorders, demonstrating unique neuroradiological features resembling those of the tubulinopathies, and support a role for DDX3X in neuronal development. Our observations further suggest a possible link between germline DDX3X variants and cancer development