The inflamed microenvironment: role on MSCs immunobiology and cancer

Inflammation and cancer are an inseparable binomial. The majority of cancers are triggered by somatic mutations and environmental factors with a common element: inflammation. Inflammation creates a microenvironment in which neoplastic cells can profit from the trophic factors secreted by inflammatory cells, useful to interfere with the anti-tumor response. Among the others, mesenchymal stem cells (MSCs) participate to microenvironment creation by a strong paracrine effect. The linkage between MSCs and inflammation is bidirectional: the inflamed microenvironment affects the complex MSCs immunobiology, but also MSCs can sustain inflammation. Here, we tried to clarify the influence of inflammation on the immunobiology of MSCs and deepen the paracrine effect of MSCs on tumor growth. MSCs were isolated from periprosthetic capsule caused by breast implant, affected by inflammation (I-MSCs). The contralateral part of the same patient, not inflamed, was used as control (C-MSCs). A panel of selected cytokines were analyzed by Real-Time PCR and ELISA. The cytokines expression was different in I-MSCs compared to C-MSCs, revealing that inflammation affects MSCs immunobiology. Then, C- and I-MSCs were indirectly co-cultured with MCF7 cells from breast adenocarcinoma. New analyses on proliferation rate and cytokines expression were performed. C- and I-MSCs gave almost the same results. The over-secretion of all the cytokines referred to the Th1 pathway and the decrease of those belonging to the Th2 pathway revealed the absence of a switch from Th1 to Th2 important to induce a chronic inflammation. The levels of TGF-β and G-CSF linked to the skill to damage the antigen-presenting cell function were decreased. In conclusion, even if MCF-7 proliferation increased after co-culture with I-MSCs, MSCs-derived paracrine effect does not sustain breast adenocarcinoma. These results absolve the breast implants from the insult to enhance adenocarcinoma onset

From nucleus pulposus mesenchymal stem cells towards neural differentiation: an interesting prospect

Regenerative medicine arouses great interest for the treatment of many neurological diseases. Since nucleus pulposus of the invertebral discs is a postembryonic vestige of the notochord, it has been hypothesized that mesenchymal stem cells (MSCs) isolated from nucleus pulposus (NP-MSCs) can more easily differentiate into neurons. In this study, MSCs from nucleus pulposus were successfully isolated and characterized. Then, neural differentiation was induced by using a medium consisted of DMEM/F12 supplemented with B27 and the growth factors FGF and EGF for 10 days. Immunocytochemistry, molecular studies, SEM and TEM microscopy analyses were performed. NP-MSCs exhibited the typical features of MSCs, revealing spindle-shape morphology, specific immunophenotype attributable to MSCs and the ability to differentiate in osteogenic and chondrogenic lineages. After neurodifferentiation induction, compared to NP-MSCs in only DMEM/F12, proliferation rate decreased and cells changed morphology acquiring an increased number of the so-called neural-like extensions. Neural progenitor marker NESTIN and mature neuronal marker ENOLASE-2 were up-regulated, while GFAP was not detected. Moreover, cells after differentiation were small rounded and fusiform, with tendency to organize in clumps; they had elongated extrusions containing oriented cytoskeletal elements, classifiable as microtubules and intermediate filaments, as visualized by SEM and TEM microscopy. Dense vesicles similar to lipid droplet were also observed. NP-MSCs in differentiation medium were able to form neurospheres. In conclusion, even if more analysis have to be done and the way to treat neurodegenerative disease with regenerative medicine is still long, NP-MSCs represent a promising resource

Sequential or Concomitant Inhibition of Cyclin-Dependent Kinase 4/6 Before mTOR Pathway in Hormone-Positive HER2 Negative Breast Cancer: Biological Insights and Clinical Implications

About 75% of all breast cancers are hormone receptor-positive (HR+). However, the efficacy of endocrine therapy is limited due to the high rate of either pre-existing or acquired resistance. In this work we reconstructed the pathways around estrogen receptor (ER), mTOR, and cyclin D in order to compare the effects of CDK4/6 and PI3K/AKT/mTOR inhibitors. A positive feedback loop links mTOR and ER that support each other. We subsequently considered whether a combined or sequential inhibition of CDK4/6 and PI3K/AKT/mTOR could ensure better results. Studies indicate that inhibition of CDK4/6 activates mTOR as an escape mechanism to ensure cell proliferation. In literature, the little evidence dealing with this topic suggests that pre-treatment with mTOR pathway inhibitors could prevent or delay the onset of CDK4/6 inhibitor resistance. Additional studies are needed in order to find biomarkers that can identify patients who will develop this resistance and in whom the sensitivity to CDK4/6 inhibitors can be restored

Crosstalk between Mesenchymal Stem Cells and tumor cells: the role of inflammation

Mesenhymal Stem Cells (MSCs) are self-renewal multipotent cells that can be isolated from different adult tissues. There is a growing interest in the role exerted by MSCs in cancer progression. MSCs exhibit a marked tropism for tumors and par- ticipate to the creation of the stroma and related inflammation, which has a critical role in carcinogenesis, progression and metastasis. Nevertheless, while many studies showed that MSCs promote tumor progression and metastasis, others reported that MSCs suppress tumor growth. These contradictory results may be due to the origin of the MSCs, their degree of differentiation, the tumor model and other factors that are not yet elucidated. Aim of this work was to establish the role of the paracrine effect exerted by MSCs isolated from inflamed (I-MSCs) and control (C-MSCs) tis- sues towards human MCF7 and KI-JK cell lines, respectively derived from a breast cancer and an anaplastic large T cell lymphoma (ALCL). After stemness characteriza- tion, MSCs were indirectly co-cultured with MCF7 or KI-JK for 7 days; subsequently the proliferation rate and the expression of specific genes were tested. Genes were selected according to their role in inflammation and cancer previously reported in literature and explicate their action by different mechanisms: chemokines with pro- (CXCL2, CXCL9) or anti-angiogenic effect (CXCL10); chemokines (CCL2, CXCL12, CXCL5) for the recruitment of myeloid-derived suppressor cells (MDSCs); interleu- kins distinctive for chronic (IL2, IL4) and acute (IL8, IL16) inflammation; cytokines belonging to the Th2 subset (CCL22, IL13, IL22, CCL17, CCL18); cytokines (IL6, IL10 and TGFβ1) involved in the manipulation of the antigen-presenting cells function. Our data confirm a role of MSCs in cancer; an increase of pro-angiogenic chemokines as well as of interleukin related to acute phase of inflammation, a general switch of the T cell response from the Th1 cell subset to the Th2 subset and the induction of MDSCs were observed. Surprisingly, these effects have been mainly found in both cancer cell lines after co-culture with C-MSCs; it may mean that I-MSCs, suffering of chronic inflammation, are less responsive than C-MSCs to new stress/stimuli. Fur- ther experiments will be necessary to better address the role of MSCs and inflamma- tion on cancer progression; nevertheless, this study highlight as MSCs are not simply guardian but active actors in cancer fate

Narrative review: predicting future molecular and clinical profiles of prostate cancer in the United States

Prostate cancer represents the most frequent tumor in men, accounting for the 21% of all diagnosed tumors, with 191,930 new cases and 33,330 deaths estimated in 2020. Advanced prostate cancer represents a heterogeneous disease, ranging from hormone naive or hormone sensitive to castration resistant. The therapeutic armamentarium for this disease has been implemented in the last years by novel hormonal therapies and chemotherapies. However, the percentage of patients who achieve complete responses still results negligible. On this scenario, the design of clinical trials investigating new therapeutic approaches represent a dramatic medical need. Predicting cancer incidence may be fundamental to design specific clinical trials, to optimize the allocation of economic resources, and to plan future cancer control programs. ERG, SPOP and DDR genes alterations can act as therapeutic targets in prostate cancer patients and can be tested to identify a gene-selected patient population to enrol in specific trials. According to our predictions, ERG gene fusions will be the most predominant molecular subtype, accounting for 69,050 new cases in 2030. Mutation in SPOP gene will be diagnosed in 16,512 tumors, corresponding to the number of cases associated with alterations in DDR genes (including 7,956 BRCA2 mutated tumors). In this article, we analyzed and discussed the future molecular and clinical profiles of prostate cancer in the United States, aimed to describe a series of distinct subpopulations and to quantify potential clinical trial candidates in the next years

The double face of cutaneous MSCs: a target to hit or a therapeutic tool in psoriasis

Le cellule staminali mesenchimali (MSCs) sono cellule staminali multipotenti. Le MSCs isolate dalla cute di pazienti psoriasici hanno caratteristiche tipiche delle lesioni, a prova di come siano strettamente coinvolte nell’insorgenza della malattia. Il presente lavoro si è focalizzato su tre aspetti correlati alle MSCs isolate da pazienti psoriasici (PSO-MSCs) non ancora noti. In primo luogo, è stata valutata l’efficacia di due inibitori del TNF-α usati nel trattamento della psoriasi sulle MSCs. È stato poi considerato il ruolo delle MSCs sane, in quanto cellule con proprietà immunomodulatorie, nel ridurre lo stato infiammatorio delle PSO-MSCs. Infine, è stata studiata la capacità delle PSO-MSCs di stimolare i cheratinociti (KCs) in senso psoriasico. Le MSCs sono state isolate dalla cute di pazienti psoriasici prima (PSO-MSCs T0) e dopo 12 settimane di terapia con anti-TNF-α (T12), e da persone sane (H-MSCs). Le cellule sono state caratterizzate secondo i criteri minimi di Dominici. L’analisi di citochine dei pathway Th1, Th2 e Th17 rivelava che le PSO-MSCs T0 avevano un profilo immunologico alterato; la terapia ristabiliva valori vicini a quelli fisiologici. Per valutare l’influenza delle H-MSCs sulle PSO-MSCs T0, si è eseguita una co-coltura indiretta. Dopo 72 ore l’iperproliferazione delle PSO-MSCs era ridotta e i livelli delle citochine riequilibrati, in maniera comparabile alle terapie prima valutate. Infine, è stato approfondito l’effetto delle PSO-MSCs sui KCs tramite co-coltura indiretta con una linea di KCs umana, le HaCaT. Le condizioni di co-coltura usate non inducevano variazione nella proliferazione cellulare; le analisi sul secretoma non chiariscono se le MSCs stimolino i KCs in senso psoriasico. In conclusione, il nostro studio prova che le terapie biologiche in uso per la psoriasi agiscono anche a livello staminale. Inoltre le H-MSCs, in quanto capaci di stimolare le PSO-MSCs verso un profilo più fisiologico, sono un promettente strumento terapeutico.Mesenchymal stem cells (MSCs) are self-renewal multipotent stem cells. MSCs isolated from skin of psoriatic patients show typical features of psoriatic lesions, confirming MSCs are strictly related to disease onset. Present work focused on three aspects related to MSCs isolated from psoriatic patients (PSO-MSCs) not yet known. Firstly, the efficacy of two TNF-α inhibitors used for psoriasis on PSO-MSCs was evaluated. Moreover, it was considered if healthy MSCs, having immunomodulatory and repair-promoting properties, can ameliorate PSO-MSCs inflammatory state. Finally, the effect of PSO-MSCs on keratinocytes (KCs) was studied. MSCs were isolated from skin of psoriatic patients before (PSO-MSCs T0) and after 12 weeks treatment with anti-TNF-α drugs (T12), and from healthy people (H-MSCs). Cells were characterized following minimal criteria of Dominici. The expression of 22 cytokines of Th1, Th2 and Th17 pathways showed that PSO-MSCs T0 had a dysregulated immunological profile but a 12 weeks therapy was able to counterweight altered levels of cytokines. To evaluate if H-MSCs, as cells with immunomodulatory properties and producers of a not-inflamed microenvironment, can positively influence non-treated PSO-MSCs, an indirect co-culture was made. A 72 hours co-culture reduced the hyperproliferation of PSO-MSCs and rebalanced cytokines levels, in a comparable way of biological therapies previously considered. Lastly, to deepen the effect of PSO-MSCs on KCs, PSO-MSCs were indirectly co-cultured with a KCs cell line, the HaCaT. Used co-culture conditions did not induce a variation in cell proliferation; even more, analyses on the secretome can not clarify whether MSCs can move KCs towards psoriasis. In conclusion, our study proves that biological treatments in use for psoriasis are effective also at the staminal level of the skin. In addition, healthy MSCs can invert the psoriatic MSCs profile towards a more physiological one, making H-MSCs a promising therapeutic tool

New miRNAs network in human mesenchymal stem cells derived from skin and amniotic fluid.

Mesenchymal stem cells (MSCs), isolated from different adult sources, have great appeal for therapeutic applications due to their simple isolation, extensive expansion potential, and high differentiative potential.In our previous studies we isolated MSCs form amniotic fluid (AF-MSCs) and skin (S-MSCs) and characterized them according to their phenotype, pluripotency, and mRNA/microRNAs (miRNAs) profiling using Card A from Life Technologies.Here, we enlarge the profiling of AF-MCSs and S-MSCs to the more recently discovered miRNAs (Card B by Life Technologies) to identify the miRNAs putative target genes and the relative signaling pathways. Card B, in fact, contains miRNAs whose role and target are not yet elucidated.The expression of the analyzed miRNAs is changing between S-MSCs and AF-MSCs, indicating that these two types of MSCs show differences potentially related to their source. Interestingly, the pathways targeted by the miRNAS deriving from Card B are the same found during the analysis of miRNAs from Card A.This result confirms the key role played by WNT and TGF-β pathways in stem cell fate, underlining as other miRNAs partially ignored up to now deserve to be reconsidered. In addition, this analysis allows including Adherens junction pathways among the mechanisms finely regulated in stem cell behavior