Study of the efficacy of new drugs in a mouse model of multiple myeloma

Il mieloma multiplo (MM) \ue8 un disordine clonale delle plasmacellule, che rappresenta approssimativamente il 10% di tutte le neoplasie ematologiche. Lo sviluppo del MM dipende dalle interazioni con il microambiente del midollo osseo (BM), che supporta la crescita, la sopravvivenza e la resistenza ai farmaci delle plasmacellule attraverso l\u2019adesione cellula-cellula e il rilascio di un grande numero di fattori di crescita, come l\u2019interleuchina-6 (IL-6) e il fattore di crescita dell\u2019endotelio vascolare (VEGF). Alcuni eventi patogenetici cruciali nel MM, come la neoangiogenesi e l\u2019osteolisi, sono indotti all\u2019interno del BM dalle stesse cellule di MM. Nonostante la chemioterapia standard sia spesso efficace nel ridurre la malattia, la remissione completa \ue8 raggiungibile solo in una minoranza di pazienti e le risposte cliniche sono raramente persistenti. Studi recenti mostrano che un numero altamente significativo di pazienti pu\uf2 ottenere buone risposte cliniche con protocolli terapeutici basati sull\u2019utilizzo di agenti anti-angiogenici o biologici, come la talidomide, lenalidomide e il bortezomib, specialmente quando utilizzati in combinazione con il desametasone. Nonostante ci\uf2, il MM rimane ancora una patologia incurabile e perci\uf2 l\u2019identificazione di nuovi obiettivi molecolari \ue8 cruciale per lo sviluppo di strategie terapeutiche innovative, che dovrebbero non solo avere effetti diretti sulle cellule neoplastiche, ma anche indiretti agendo sul microambiente midollare che supporta il MM. A questo scopo, l\u2019utilizzo di modelli murini con un microambiente di sviluppo della malattia molto simile a quello umano risulta utile per prevedere gli effetti clinici nei pazienti. Tra questi, il 5TMM \ue8 di particolare interesse, in quanto origina da un MM sviluppato spontaneamente in topi anziani del ceppo C57BL/KalwRij. Le caratteristiche di tale modello, che comprendono la localizzazione midollare delle cellule di MM, la presenza della componente monoclonale nel siero, l\u2019induzione di osteolisi e la neoangiogenesi nel midollo, lo rendono molto simile al MM umano. In questo studio, sono stati esaminati gli effetti in vitro di diversi nuovi farmaci in fase di sviluppo della Ditta Novartis su linee cellulari umane e murine e sono state testate queste molecole in vivo nel modello 5T33MM. La molecola T8 induce una riduzione dell\u2019attivit\ue0 cellulare solo per alte concentrazioni, con un valore di IC50 di 4315 nM dopo 48 ore di incubazione; per questa ragione non \ue8 stato utilizzato negli studi successivi. Come atteso, la molecola N80 non mostra alcuna attivit\ue0 citotossica sulle cellule trattate rispetto ai controlli. Comunque, quando \ue8 stato testata in vivo per un trattamento preventivo e curativo, \ue8 stata dimostrata un\u2019immediata tossicit\ue0 e la riduzione del carico tumorale. Non si evidenziano differenze nei livelli di IgG2b monoclonale nel siero dei topi trattati con N80 utilizzando il protocollo terapeutico rispetto ai controlli, mentre il protocollo preventivo induce una riduzione statisticamente significativa nei livelli di IgG2b. L9, N0 e N2 sono in grado d\u2019inibire l\u2019attivit\ue0 delle cellule di MM in vitro. Si sono dimostrati i composti pi\uf9 potenti, con valori nanomolari di IC50 e in grado di indurre una riduzione della vitalit\ue0 cellulare, correlata con l\u2019apoptosi cellulare in vitro. Queste molecole riducono la produzione della componente monoclonale e i livelli sierici di VEGF in vivo. Quindi, queste molecole sono in grado di ridurre il carico tumorale, sebbene non modifichino la sopravvivenza dei topi nel modello 5T33vtMM con gli attuali protocolli terapeutici utilizzati.Multiple myeloma (MM) is a clonal plasma cell disorder accounting for approximately 10% of all haematological malignancies. MM development dramatically depends on plasma cell interactions with bone marrow (BM) microenvironment, which supports plasma cell growth, survival and drug resistance through cell-cell adhesion and release of a large number of growth factors, including interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF). Some crucial pathogenetic events in MM, such as neoangiogenesis and osteolysis, are ignited inside BM by MM cells themselves. Although standard chemotherapy is usually effective in lowering the disease burden, complete remission is achievable only in a minority of patients, and clinical responses are rarely persistent. Recent studies showed that a significantly higher number of patients may achieve clinical major responses when treated with therapeutic schedules based on the use of anti-angiogenic or biological agents, such as thalidomide, lenalidomide and bortezomib, especially when employed in combination with dexamethasone. However, MM still remains an incurable disease. Therefore, the identification of new key targets is crucial for the development of innovative therapeutic strategies, which should have not only direct effects on MM cells, but also interfering effects on MM-supporting BM microenvironment. To this aim, the use of mouse models that may closely resemble human MM development are mostly useful to foresee the clinical effects in patients. Some kinds of murine MM, such as 5TMM, originate from spontaneously developed MM in elderly mice of C57BL/KalwRij strain. The characteristics of these models, including the localization of the MM cells in the BM, the presence of serum M-component, the induction of osteolytic bone disease and neo-angiogenesis in the BM, are similar to human MM. In this study, we examined the effects of several new drugs in different murine and human myeloma cell lines and we tested these molecules in vivo in the murine 5T33MM model. T8 induced a reduction in cell activity only for high concentrations of molecule, with IC50 value of 4315 nM after 48 hours of incubation; for this reason it was not used for further studies. As expected, N80 did not show any cytotoxic activity on treated cells as compared to controls. However, when tested in vivo for a preventive and curative treatment, N80 showed immediate toxicity and reduction of the tumor burden. There were not differences in IgG2b levels in serum of mice treated with N80 using the therapeutical schedule as compared to controls, while the preventive schedule induced a statistically significant reduction in IgG2b levels. L9, N0 and N2 were able to inhibit the activity of MM cells in vitro. They appeared the most potent compounds, with nanomolar IC50 values, capable of reducing cell viability, in correlation with cell apoptosis in vitro. These molecules reduced the M-component and VEGF serum levels in vivo and eventually the tumor burden, although they did not modify significantly, with the treatment schedules employed, the overall survival in 5T33vtMM model

Endothelin-1 receptor blockade as new possible therapeutic approach in multiple myeloma.

New effective treatments are needed to improve outcomes for multiple myeloma (MM) patients. Receptors with restricted expression on plasmacells (PCs) represent attractive new therapeutic targets. The endothelin-1(EDN1) axis, consisting of EDN1 acting through EDN-receptor A(EDNRA) and B (EDNRB), was previously shown to be overexpressed inseveral tumours, including MM. However, there is incomplete understand-ing of how EDN1 axis regulates MM growth and response to therapy.Besides EDNRA, the majority of MM cell lines and primary malignant PCsexpress high levels of EDNRB and release EDN1. Similarly, bone-marrowmicroenvironment cells also secrete EDN1. Investigating the extent of epi-genetic dysregulation of EDNRB gene in MM, we found that hypermethyla-tion of EDNRB promoter and subsequent down-regulation of EDNRB genewas observed in PCs or B lymphocytes from healthy donors compared toEDNRB-expressing malignant PCs. Pharm acological blockade with the dualEDN1 receptor antagonist bosentan decreased cell viability and MAPK acti-vation of U266 and RPMI-8226 cells. Interestingly, the combination ofbosentan and the proteasome inhibitor bortezomib, currently approved forMM treatment, resulted in synergistic cytotoxic effects. Overall, our datahas uncovered EDN1-mediated autocrine and paracrine mechanisms thatregulate malignant PCs growth and drug response, and support EDN1receptors as new therapeutic targets in MM

SPOP Deregulation Improves the Radiation Response of Prostate Cancer Models by Impairing DNA Damage Repair

Speckle-type POZ (pox virus and zinc finger protein) protein (SPOP) is the most commonly mutated gene in prostate cancer (PCa). Recent evidence reports a role of SPOP in DNA damage response (DDR), indicating a possible impact of SPOP deregulation on PCa radiosensitivity. This study aimed to define the role of SPOP deregulation (by gene mutation or knockdown) as a radiosensitizing factor in PCa preclinical models. To express WT or mutant (Y87N, K129E and F133V) SPOP, DU145 and PC-3 cells were transfected with pMCV6 vectors. Sensitivity profiles were assessed using clonogenic assay and immunofluorescent staining of γH2AX and RAD51 foci. SCID xenografts were treated with 5 Gy single dose irradiation using an image-guided small animal irradiator. siRNA and miRNA mimics were used to silence SPOP or express the SPOP negative regulator miR-145, respectively. SPOP deregulation, by either gene mutation or knockdown, consistently enhanced the radiation response of PCa models by impairing DDR, as indicated by transcriptome analysis and functionally confirmed by decreased RAD51 foci. SPOP silencing also resulted in a significant downregulation of RAD51 and CHK1 expression, consistent with the impairment of homologous recombination. Our results indicate that SPOP deregulation plays a radiosensitizing role in PCa by impairing DDR via downregulation of RAD51 and CHK1. View Full-Tex

Efficacy assessment of interferon-alpha-engineered mesenchymal stromal cells in a mouse plasmacytoma model

Bone marrow mesenchymal stromal cells (BM-MSCs) may survive and proliferate in the presence of cycling neoplastic cells. Exogenously administered MSCs are actively incorporated in the tumor as stromal fibroblasts, thus competing with the local mesenchymal cell precursors. For this reason, MSCs have been suggested as a suitable carrier for gene therapy strategies, as they can be genetically engineered with genes encoding for biologically active molecules, which can inhibit tumor cell proliferation and enhance the anti-tumor immune response. We used BM-MSCs engineered with the murine interferon-alpha (IFN-alpha) gene (BM-MSCs/IFN-alpha) to assess in a mouse plasmacytoma model the efficacy of this approach towards neoplastic plasma cells. We found that IFN-alpha can be efficiently produced and delivered inside the tumor microenvironment. Subcutaneous multiple administration of BM-MSCs/IFN-alpha significantly hampered the tumor growth in vivo and prolonged the overall survival of mice. The anti-tumor effect was associated with enhanced apoptosis of tumor cells, reduction in microvessel density, and ischemic necrosis. By contrast, intravenous administration of BM-MSCs/IFN-alpha did not significantly modify the survival of mice, mainly as a consequence of an excessive entrapment of injected cells in the pulmonary vessels. In conclusion, BM-MSCs/IFN-alpha are effective in inhibiting neoplastic plasma cell growth; however, systemic administration of engineered MSCs still needs to be improved to make this approach potentially suitable for the treatment of multiple myeloma

Characteristics and patterns of care of endometrial cancer before and during COVID-19 pandemic

Objective: Coronavirus disease 2019 (COVID-19) outbreak has correlated with the disruption of screening activities and diagnostic assessments. Endometrial cancer (EC) is one of the most common gynecological malignancies and it is often detected at an early stage, because it frequently produces symptoms. Here, we aim to investigate the impact of COVID-19 outbreak on patterns of presentation and treatment of EC patients. Methods: This is a retrospective study involving 54 centers in Italy. We evaluated patterns of presentation and treatment of EC patients before (period 1: March 1, 2019 to February 29, 2020) and during (period 2: April 1, 2020 to March 31, 2021) the COVID-19 outbreak. Results: Medical records of 5,164 EC patients have been retrieved: 2,718 and 2,446 women treated in period 1 and period 2, respectively. Surgery was the mainstay of treatment in both periods (p=0.356). Nodal assessment was omitted in 689 (27.3%) and 484 (21.2%) patients treated in period 1 and 2, respectively (p<0.001). While, the prevalence of patients undergoing sentinel node mapping (with or without backup lymphadenectomy) has increased during the COVID-19 pandemic (46.7% in period 1 vs. 52.8% in period 2; p<0.001). Overall, 1,280 (50.4%) and 1,021 (44.7%) patients had no adjuvant therapy in period 1 and 2, respectively (p<0.001). Adjuvant therapy use has increased during COVID-19 pandemic (p<0.001). Conclusion: Our data suggest that the COVID-19 pandemic had a significant impact on the characteristics and patterns of care of EC patients. These findings highlight the need to implement healthcare services during the pandemic

Practice patterns and 90-day treatment-related morbidity in early-stage cervical cancer

To evaluate the impact of the Laparoscopic Approach to Cervical Cancer (LACC) Trial on patterns of care and surgery-related morbidity in early-stage cervical cancer

Absence of lingual frenulum in children with Ehlers-Danlos Syndrome: a retrospective study of forty cases and literature review of a twenty years long debate

Ehlers-Danlos syndrome (EDS) is part of connective tissue disorders and is characterized by skin hyperextensibility, joint hypermobility, easy bruising and other severe manifestations such as epilepsy, pneumothorax, arterial rupture and bowel perforation. In 2017 a new classification was published, indicating major and minor criteria for each form of EDS. Further reports in the past years tried to determine whether or not the absence of lingual frenulum should be included in minor criteria for the diagnosis of EDS, but a consensus has still not been reached. The aim of this study was to assess the clinical relevance of lingual frenulum absence, evaluating its prevalence in a cohort of EDS pediatric patients and comparing it to a group of controls

The contribution of intraepithelial inflammatory cells to the histological diagnosis of microscopic esophagitis

Background: Diagnosis of gastro-esophageal reflux disease (GERD) is mainly based on symptom evaluation, possibly coupled with various invasive investigations (endoscopy, pH-metry, and impedance). New input has recently come from histology: in this context, the diagnostic role of inflammatory cells other than eosinophils and neutrophils is still debated. Aim of the study is to evaluate the diagnostic relevance of intraepithelial lymphocytes, mast cells, and Langerhans cells in GERD-associated microscopic esophagitis (GAME). Methods: Twenty healthy volunteers and 119 patients with GERD symptoms were prospectively recruited and subdivided, on the basis of endoscopy and pH-metry, in erosive (ERD, 48) and non-erosive disease (NERD, 71). Biopsy samples at 2 cm above the Z-line and at Z-line were evaluated for GERD-associated histological lesions (basal cell hyperplasia, papillae elongation, intercellular space dilatation, intraepithelial eosinophils, and neutrophils). Immunohistochemistry for T lymphocytes (CD3, CD4, CD8), B lymphocytes (CD20), NK cells (CD56), macrophages (CD68), mast cells (c-Kit), and Langerhans cells (S100) was performed. Results: Among inflammatory cells, only intraepithelial T lymphocytes (ITLs) showed statistical correlation with the other histological lesions both in ERD and NERD. ITLs distinguished GERD patients from controls with good sensitivity and specificity (85.5 and 85 % at 2 cm above Z-line; 89.5 and 75 % at Z-line) when a cut-off of 20 cells was applied. An analysis of the T subpopulations found a CD4+/CD8+ ratio close to 1:1; B cells, mast cells, Langerhans cells, NK cells, and macrophages showed a limited role in GERD. Conclusions: ITL evaluation represents an additional useful parameter in the histological evaluation of GAME

Fibroid Removal after Myomectomy: An Overview on the Problems of Power Morcellation

The authors reviewed uterine fibroid (UF) morcellation and its potential consequences, notably a hypothetical spread and dissemination of occult uterine leiomyosarcoma (LMS) tissue, evaluating the effect of laparoscopic versus open myomectomies with and without morcellation on patients’ outcomes, as well as related medical-legal issues. MEDLINE and PubMed search was performed for the years 1990–2021, using a combination of keywords on this topic. Relevant articles were identified and included in this narrative review. There is an individual risk, for all patients, for LMS diagnosis after myomectomy. However, the risk for occult LMS diagnosis during a laparoscopic myomectomy is generally reduced when the guidelines of scientific societies are followed, with an overall benefit from the laparoscopic approach with morcellation in appropriate cases. Gynecological societies do not ban morcellation and laparoscopic hysterectomy/myomectomy per se, but recommend their use on the basis of the patients’ clinical characteristics. It is suggested for gynecologists to provide detailed information to patients when obtaining an informed consent for open or laparoscopic hysterectomy/myomectomy. A detailed preoperative assessment of patients and the risk benefit ratio of laparoscopic morcellation of uterine mass could overcome the “a priori” banning of the morcellation technique

In Vitro and In Vivo Model of EBV-Positive Non-Hodgkin Plasmablastic Lymphoma with Focal Plasmacytic Differentiation

Assessment of EBV-positive large B-cell lympoma cell line with plasmacytic differentiatio