Bladder cancer is lethal in its advanced, muscle-invasive phase with very limited therapeutic advances
. Recent molecular characterization has defined new (epi)genetic drivers and potential targets ...for bladder cancer
. The immune checkpoint inhibitors have shown remarkable efficacy but only in a limited fraction of bladder cancer patients
. Here, we show that high G9a (EHMT2) expression is associated with poor clinical outcome in bladder cancer and that targeting G9a/DNMT methyltransferase activity with a novel inhibitor (CM-272) induces apoptosis and immunogenic cell death. Using an immunocompetent quadruple-knockout (Pten
; Trp53
; Rb1
; Rbl1
) transgenic mouse model of aggressive metastatic, muscle-invasive bladder cancer, we demonstrate that CM-272 + cisplatin treatment results in statistically significant regression of established tumors and metastases. The antitumor effect is significantly improved when CM-272 is combined with anti-programmed cell death ligand 1, even in the absence of cisplatin. These effects are associated with an endogenous antitumor immune response and immunogenic cell death with the conversion of a cold immune tumor into a hot tumor. Finally, increased G9a expression was associated with resistance to programmed cell death protein 1 inhibition in a cohort of patients with bladder cancer. In summary, these findings support new and promising opportunities for the treatment of bladder cancer using a combination of epigenetic inhibitors and immune checkpoint blockade.
Bladder cancer is a current clinical and social problem. At diagnosis, most patients present with nonmuscle-invasive tumors, characterized by a high recurrence rate, which could progress to ...muscle-invasive disease and metastasis. Bone morphogenetic protein (BMP)-dependent signaling arising from stromal bladder tissue mediates urothelial homeostasis by promoting urothelial cell differentiation. However, the possible role of BMP ligands in bladder cancer is still unclear.
Tumor and normal tissue from 68 patients with urothelial cancer were prospectively collected and analyzed for expression of BMP and macrophage markers. The mechanism of action was assessed
by experiments with bladder cancer cell lines and peripheral blood monocyte-derived macrophages.
We observed
expression is associated and favored type II macrophage differentiation.
experiments showed that both recombinant BMP4 and BMP4-containing conditioned media from bladder cancer cell lines favored monocyte/macrophage polarization toward M2 phenotype macrophages, as shown by the expression and secretion of IL10. Using a series of human bladder cancer patient samples, we also observed increased expression of
in advanced and undifferentiated tumors in close correlation with epithelial-mesenchymal transition (EMT). However, the p-Smad 1,5,8 staining in tumors showing EMT signs was reduced, due to the increased miR-21 expression leading to reduced
expression.
These findings suggest that BMP4 secretion by bladder cancer cells provides the M2 signal necessary for a protumoral immune environment. In addition, the repression of
by miR-21 makes the tumor cells refractory to the prodifferentiating actions mediated by BMP ligands, favoring tumor growth.
.
Different strategies are being investigated for treating PMM2‐CDG, the most common congenital disorder of glycosylation. The use of pharmacochaperones (PCs) is one of the most promising. The present ...work characterizes the expression, stability, and enzymatic properties of 15 previously described clinical variants of the PMM2 protein, four novel variants, the Pmm2 mouse variant p.Phe115Leu, and its p.Phe119Leu human counterpart, with the aim of extending the potential use of pharmacochaperoning treatment. PMM2 variants were purified as stable homodimers, except for p.Asp65Gly, p.Ile120Thr, and p.Thr237Lys (no expression detected), p.Thr226Ser and p.Val231Met (aggregates), and p.Glu93Ala, p.Phe119Leu, and p.Phe115Leu (partial dissociated). Enzyme activity analyses identified severe variants and milder ones. Pure dimeric mutant proteins showed a reduction in thermal stability except for p.Asn216Asp. The thermal stability of all the unstable mutants was recovered in the presence of the PC compound VIII. This study adds to the list of destabilizing human variants amenable to rescue by small chemical compounds that increase the stability/activity of PMM2. The proposed platform can be reliably used for assessing the disease‐causing effects of PMM2 missense variants, for assessing the correlation between genotype and phenotype, for confirming new clinical defects, and for identifying destabilizing mutations amenable to rescue by PCs.
Bladder Cancer (BC) represents a clinical and social challenge due to its high incidence and recurrence rates, as well as the limited advances in effective disease management. Currently, a ...combination of cytology and cystoscopy is the routinely used methodology for diagnosis, prognosis and disease surveillance. However, both the poor sensitivity of cytology tests as well as the high invasiveness and big variation in tumour stage and grade interpretation using cystoscopy, emphasizes the urgent need for improvements in BC clinical guidance. Liquid biopsy represents a new non-invasive approach that has been extensively studied over the last decade and holds great promise. Even though its clinical use is still compromised, multiple studies have recently focused on the potential application of biomarkers in liquid biopsies for BC, including circulating tumour cells and DNA, RNAs, proteins and peptides, metabolites and extracellular vesicles. In this review, we summarize the present knowledge on the different types of biomarkers, their potential use in liquid biopsy and clinical applications in BC.
The protein content of dry biomass of the microalgae
Porphyridium cruentum,
Scenedesmus almeriensis, and
Muriellopsis sp
. and of the cyanobacteria
Synechocystis aquatilis and
Arthrospira platensis ...was measured by the Lowry method following disruption of the cells by milling with inert ceramic particles. The measurements were compared with the Kjeldahl method and by elemental analysis. The nitrogen-to-protein conversion factors for biomass obtained from exponentially growing cells with a steady state doubling time of ∼23
h were 5.95 for nitrogen measured by Kjeldahl and 4.44 for total nitrogen measured by elemental analysis. The protein content in dry biomass ranged from 30% to 55%. The above conversion factors are useful for estimating the protein content of microalgal biomass produced in rapid steady state growth as encountered in many commercial production processes.
We investigate the relation between the maximum cardinality N of the level sets of a Lipschitz quotient mapping of the plane and the ratio between its Lipschitz and co‐Lipschitz constants, with ...respect to the polygonal norms, and establish that bounds of 1/N previously shown to be sharp for Euclidean norm stay sharp for polygonal n‐norms if and only if n is not divisible by 4.
In this study, we have developed a metal‐mediated synthetic method for incorporating the 1,1,2,2‐tetrafluoroethyl (CF2CF2H) motif into unactivated, electron‐rich alkenyl iodides using cross‐coupling ...reactions. We discovered that the stable Cu(CF2CF2H)(PPh2Me)3 complex, while unreactive with these substrates, serves as a P‐ligated reservoir for the formation of solvent‐stabilized “ligandless” and reactive CuCF2CF2H species. This transformation occurs upon addition of CuBr, which partially scavenges the P‐ligand in the form of CuBr(PPh2Me)3. The resulting in situ generated solvent stabilized “ligandless” system significantly enhances the reactivity of the complex, particularly towards challenging electron‐rich substrates. This advancement enables the synthesis of HCF2CF2‐glycals, as well as nucleosides/nucleobases and arenes.
Bladder cancer is a clinical and social problem due to its high incidence and recurrence rates. It frequently appears in elderly patients showing other medical comorbidities that hamper the use of ...standard chemotherapy. We evaluated the activity of CDK4/6 inhibitor as a new therapy for patients unfit for cisplatin (CDDP).
Bladder cancer cell lines were tested for
sensitivity to CDK4/6 inhibition. A novel metastatic bladder cancer mouse model was developed and used to test its
activity.
Cell lines tested were sensitive to CDK4/6 inhibition, independent on
gene status. Transcriptome analyses and knockdown experiments revealed a major role for FOXM1 in this response. CDK4/6 inhibition resulted in reduced FOXM1 phosphorylation
and
and showed synergy with CDDP, allowing a significant tumor regression. FOXM1 exerted important oncogenic roles in bladder cancer.
CDK4/6 inhibitors, alone or in combination, are a novel therapeutic strategy for patients with advanced bladder cancer previously classified as unfit for current treatment options.
Investigating the evolution of human speech is difficult and controversial because human speech surpasses nonhuman primate vocal communication in scope and flexibility 1–3. Monkey vocalizations have ...been assumed to be largely innate, highly affective, and stereotyped for over 50 years 4, 5. Recently, this perception has dramatically changed. Current studies have revealed distinct learning mechanisms during vocal development 6–8 and vocal flexibility, allowing monkeys to cognitively control when 9, 10, where 11, and what to vocalize 10, 12, 13. However, specific call features (e.g., duration, frequency) remain surprisingly robust and stable in adult monkeys, resulting in rather stereotyped and discrete call patterns 14. Additionally, monkeys seem to be unable to modulate their acoustic call structure under reinforced conditions beyond natural constraints 15, 16. Behavioral experiments have shown that monkeys can stop sequences of calls immediately after acoustic perturbation but cannot interrupt ongoing vocalizations, suggesting that calls consist of single impartible pulses 17, 18. Using acoustic perturbation triggered by the vocal behavior itself and quantitative measures of resulting vocal adjustments, we show that marmoset monkeys are capable of producing calls with durations beyond the natural boundaries of their repertoire by interrupting ongoing vocalizations rapidly after perturbation onset. Our results indicate that marmosets are capable of interrupting vocalizations only at periodic time points throughout calls, further supported by the occurrence of periodically segmented phees. These ideas overturn decades-old concepts on primate vocal pattern generation, indicating that vocalizations do not consist of one discrete call pattern but are built of many sequentially uttered units, like human speech.
•Marmosets interrupt ongoing vocalizations rapidly after acoustic perturbation•Interruptions happen at periodic time points, indicating precisely tuned call units•Segmented phee calls have rapid, precise elements matching these time points•Calls are built from many serially uttered units rather than one discrete pattern
Pomberger et al. show that marmoset monkey calls do not consist of one discrete call pattern but are built out of many sequentially uttered units, like human speech. These findings explain the monkeys’ capability to interrupt their calls only at periodic time points within calls and are supported by the occurrence of periodically segmented calls.
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