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Abstract Androgen deprivation therapy (ADT) targeting androgen/androgen receptor (AR)- signaling pathways is the main therapy for advanced prostate cancer (PCa). However, ADT eventually fails in most patients who consequently develop castration-resistant prostate cancer (CRPC). While more potent AR antagonists and blockers for androgen synthesis were developed to improve clinical outcomes, they also show to induce more diverse CRPC phenotypes. Specifically, the AR- and neuroendocrine-null PCa, DNPC, occurs in abiraterone and enzalutamide-treated patients. Here, we uncover that current ADT induces aberrant HGF/MET signaling activation that further elevates Wnt/β-catenin signaling in human DNPC samples. Co-activation of HGF/MET and Wnt/β-catenin axes in mouse prostates induces DNPC-like lesions. Single-cell RNA sequencing analyses identify increased expression and activity of XPO1 and ribosomal proteins in mouse DNPC-like cells. Elevated expression of XPO1 and ribosomal proteins is also identified in clinical DNPC specimens. Inhibition of XPO1 and ribosomal pathways represses DNPC growth in both in vivo and ex vivo conditions, evidencing future therapeutic targets.

IntroductionDespite aggressive standard-of-care therapy, including surgery, radiation, and chemotherapy, glioblastoma recurrence is almost inevitable and uniformly lethal. Activation of glioma-intrinsic Wnt/β-catenin signaling is associated with a poor prognosis and the proliferation of glioma stem-like cells, leading to malignant transformation and tumor progression. Impressive results in a subset of cancers have been obtained using immunotherapies including anti-CTLA4, anti-PD-1, and anti-PD-L1 or chimeric antigen receptor (CAR) T cell therapies. However, the heterogeneity of tumors, low mutational burden, single antigen targeting, and associated antigen escape contribute to non-responsiveness and potential tumor recurrence despite these therapeutic efforts. In the current study, we determined the effects of the small molecule, highly specific Wnt/CBP (CREB Binding Protein)/β-catenin antagonist ICG-001, on glioma tumor cells and the tumor microenvironment (TME)–including its effect on immune cell infiltration, blood vessel decompression, and metabolic changes.MethodsUsing multiple glioma patient-derived xenografts cell lines and murine tumors (GL261, K-Luc), we demonstrated in vitro cytostatic effects and a switch from proliferation to differentiation after treatment with ICG-001.ResultsIn these glioma cell lines, we further demonstrated that ICG-001 downregulated the CBP/β-catenin target gene Survivin/BIRC5–a hallmark of Wnt/CBP/β-catenin inhibition. We found that in a syngeneic mouse model of glioma (K-luc), ICG-001 treatment enhanced tumor infiltration by CD3+ and CD8+ cells with increased expression of the vascular endothelial marker CD31 (PECAM-1). We also observed differential gene expression and induced immune cell infiltration in tumors pretreated with ICG-001 and then treated with CAR T cells as compared with single treatment groups or when ICG-001 treatment was administered after CAR T cell therapy.DiscussionWe conclude that specific Wnt/CBP/β-catenin antagonism results in pleotropic changes in the glioma TME, including glioma stem cell differentiation, modulation of the stroma, and immune cell activation and recruitment, thereby suggesting a possible role for enhancing immunotherapy in glioma patients.

The therapeutic potential of targeting the β-catenin/CBP interaction has been demonstrated in a variety of preclinical tumor models with a small molecule inhibitor, ICG-001, characterized as a β-catenin/CBP antagonist. Despite the high binding specificity of ICG-001 for the N-terminus of CBP, this β-catenin/CBP antagonist exhibits pleiotropic effects. Our recent studies found global changes in three-dimensional (3D) chromatin architecture in response to disruption of the β-catenin/CBP interaction in pancreatic cancer cells. However, an understanding of how the functional crosstalk between the antagonist and the β-catenin/CBP interaction affects changes in 3D chromatin architecture and, thereby, gene expression and downstream effects remains to be elucidated. Here, we perform Hi-C analyses on canonical and patient-derived pancreatic cancer cells before and after treatment with ICG-001. In addition to global alteration of 3D chromatin domains, we unexpectedly identify insulin signaling genes enriched in the altered chromatin domains. We further demonstrate that the chromatin loops associated with insulin signaling genes are significantly weakened after ICG-001 treatment. We finally elicit the deletion of a looping of IRS1—a key insulin signaling gene—significantly impeding pancreatic cancer cell growth, indicating that looping-mediated insulin signaling might act as an oncogenic pathway to promote pancreatic cancer progression. Our work shows that targeting aberrant insulin chromatin looping in pancreatic cancer might provide a therapeutic benefit.

Abstract In oral squamous cell carcinoma (OSCC), a highly aggressive and frequently lethal malignancy, the role and action mechanism of the microtubule regulatory protein CDK5RAP2 have not been fully understood. Here, we show that CDK5RAP2 is highly expressed in OSCC and its expression correlates with clinical stage and lymph node metastasis of the disease. The expression of CDK5RAP2 is regulated by the Wnt signaling pathway. Depletion of CDK5RAP2 inhibits the tumorigenesis and migration of OSCC cells and alters the OSCC cancer stem (-like) cell (CSC) signature. Notably, suppression of CDK5RAP2 expression disrupts spindle orientation during mitosis. Collectively, these results identify CDK5RAP2 as a potential CSC marker and reveal a mechanism that controls the CSC population in OSCC.

A 66-year-old male with a history of low back pain was found to have discitis and osteomyelitis. Biopsy and PCR testing revealed Streptococcus cristatus infection. This bacteria does not typically cause disease, and only a few cases in the literature have reported it to cause infection in the bones or joints. This case illustrates that vertebral osteomyelitis with a rare causative agent, S. cristatus, is possible and can be identified with PCR. Treatment typically requires long-term antibiotics tailored to the causative agent for a minimum of 6 weeks and can sometimes include surgical management.

A deeper understanding of the biology of therapy resistance is important for the development of optimal strategies to attain complete cancer cures [...]

Nasopharyngeal carcinoma (NPC) is an Epstein–Barr virus (EBV)-associated malignancy ranking as the 23rd most common cancer globally, while its incidence rate ranked the 9th in southeast Asia. Tumor metastasis is the dominant cause for treatment failure in NPC and metastatic NPC is yet incurable. The Wnt/β-catenin signaling pathway plays an important role in many processes such as cell proliferation, differentiation, epithelial–mesenchymal transition (EMT), and self-renewal of stem cells and cancer stem cells (CSCs). Both the EMT process and CSCs are believed to play a critical role in cancer metastasis. We here investigated whether the specific CBP/β-catenin Wnt antagonist, IGC-001, affects the metastasis of NPC cells. We found that ICG-001 treatment could reduce the adhesion capability of NPC cells to extracellular matrix and to capillary endothelial cells and reduce the tumor cell migration and invasion, events which are closely associated with distant metastasis. Through a screening of EMT and CSC-related microRNAs, it was found that miR-134 was consistently upregulated by ICG-001 treatment in NPC cells. Very few reports have mentioned the functional role of miR-134 in NPC, except that the expression was found to be downregulated in NPC. Transient transfection of miR-134 into NPC cells reduced their cell adhesion, migration, and invasion capability, but did not affect the growth of CSC-enriched tumor spheres. Subsequently, we found that the ICG-001-induced miR-134 expression resulting in downregulation of integrin β1 (ITGB1). Such downregulation reduced cell adhesion and migration capability, as demonstrated by siRNA-mediated knockdown of ITGB1. Direct targeting of ITGB1 by miR-134 was confirmed by the 3′-UTR luciferase assay. Lastly, using an in vivo lung metastasis assay, we showed that ICG-001 transient overexpression of miR-134 or stable overexpression of miR-134 could significantly reduce the lung metastasis of NPC cells. Taken together, we present here evidence that modulation of Wnt/β-catenin signaling pathway could inhibit the metastasis of NPC through the miR-134/ITGB1 axis.

Cancer cell phenotypes evolve during a tumor's treatment. In some cases, tumor cells acquire cancer stem cell-like (CSL) traits such as resistance to chemotherapy and diminished differentiation; therefore, targeting these cells may be therapeutically beneficial. In this study we show that in progressive estrogen receptor positive (ER+) metastatic breast cancer tumors, resistant subclones that emerge following chemotherapy have increased CSL abundance. Further, in vitro organoid growth of ER+ patient cancer cells also shows that chemotherapy treatment leads to increased abundance of ALDH+/CD44+ CSL cells. Chemotherapy induced CSL abundance is blocked by treatment with a pan-HDAC inhibitor, belinostat. Belinostat treatment diminished both mammosphere formation and size following chemotherapy, indicating a decrease in progenitor CSL traits. HDAC inhibitors specific to class IIa (HDAC4, HDAC5) and IIb (HDAC6) were shown to primarily reverse the chemo-resistant CSL state. Single-cell RNA sequencing analysis with patient samples showed that HDAC targets and MYC signaling were promoted by chemotherapy and inhibited upon HDAC inhibitor treatment. In summary, HDAC inhibition can block chemotherapy-induced drug resistant phenotypes with ‘one-two punch’ strategy in refractory breast cancer cells.

Objective Mobile health platforms have become an important component of pain self-management programs and hundreds of mobile applications are commercially available for patients to monitor pain. However, few of these applications have been developed in collaboration with healthcare professionals or have been critically evaluated. Manage My Pain is a user-driven mobile health platform developed by ManagingLife in collaboration with clinician researchers. Manage My Pain allows patients to keep a “pain record” and supports communication of this information with clinicians. The current report describes a user engagement study of Manage My Pain among patients at the Transitional Pain Service (TPS) at Toronto General Hospital, a multidisciplinary clinic for patients at high risk of developing postsurgical pain. Methods Patients at the TPS were encouraged to register on Manage My Pain as one component of a larger, non-randomized prospective study of treatment predictors and treatment enhancement. Uptake of the application and rates of registration, use, and retention were tracked for 90 days. Results Of the 196 patients who consented to the larger study, 132 (67%) also provided consent to the Manage My Pain component, indicating that they found this to be an acceptable treatment adjunct, and 119 (61%) completed registration. Of those who used the app, 67.9% and 43.2% continued to use Manage My Pain beyond 30 and 90 days, respectively. On average, users engaged with the app for 93.14 days (SD = 151.9 days) logged an average of 47.39 total records (SD = 136.1). Conclusions Manage My Pain was found acceptable by a majority of patients at an academic pain management program. Rates of user registration and retention were favorable compared to those reported by other applications. Further research is needed to develop strategies to retain users and maximize patient benefit.

The integration of cellular status with metabolism is critically important and the coupling of energy production and cellular function is highly evolutionarily conserved. This has been demonstrated in stem cell biology, organismal, cellular and tissue differentiation and in immune cell biology. However, a molecular mechanism delineating how cells coordinate and couple metabolism with transcription as they navigate quiescence, growth, proliferation, differentiation and migration remains in its infancy. The extreme N-termini of the Kat3 coactivator family members, CBP and p300, by far the least homologous regions with only 66% identity, interact with members of the nuclear receptor family, interferon activated Stat1 and transcriptionally competent β-catenin, a critical component of the Wnt signaling pathway. We now wish to report based on multiomic and functional investigations, utilizing p300 knockdown, N-terminal p300 edited and p300 S89A edited cell lines and p300 S89A knockin mice, that the N-termini of the Kat3 coactivators provide a highly evolutionarily conserved hub to integrate multiple signaling cascades to coordinate cellular metabolism with the regulation of cellular status and function.

Differential usage of Kat3 coactivators, CBP and p300, by β-catenin is a fundamental regulatory mechanism in stem cell maintenance and initiation of differentiation and repair. Based upon our earlier pharmacologic studies, p300 serine 89 (S89) is critical for controlling differential coactivator usage by β-catenin via post-translational phosphorylation in stem/progenitor populations, and appears to be a target for a number of kinase cascades. To further investigate mechanisms of signal integration effected by this domain, we generated p300 S89A knock-in mice. We show that S89A mice are extremely sensitive to intestinal insult resulting in colitis, which is known to significantly increase the risk of developing colorectal cancer. We demonstrate cell intrinsic differences, and microbiome compositional differences and differential immune responses, in intestine of S89A versus wild type mice. Genomic and proteomic analyses reveal pathway differences, including lipid metabolism, oxidative stress response, mitochondrial function and oxidative phosphorylation. The diverse effects on fundamental processes including epithelial differentiation, metabolism, immune response and microbiome colonization, all brought about by a single amino acid modification S89A, highlights the critical role of this region in p300 as a signaling nexus and the rationale for conservation of this residue and surrounding region for hundreds of million years of vertebrate evolution.

The Wnt signaling pathway is one of the major signaling pathways used by cancer stem cells (CSC). Ecotropic Viral Integration Site 1 (EVI1) has recently been shown to regulate oncogenic development of tumor cells by interacting with multiple signaling pathways, including the Wnt signaling. In the present study, we found that the Wnt modulator ICG-001 could inhibit the expression of EVI1 in nasopharyngeal carcinoma (NPC) cells. Results from loss-of-function and gain-of-function studies revealed that EVI1 expression positively regulated both NPC cell migration and growth of CSC-enriched tumor spheres. Subsequent studies indicated ICG-001 inhibited EVI1 expression via upregulated expression of miR-96. Results from EVI1 3′UTR luciferase reporter assay confirmed that EVI1 is a direct target of miR-96. Further mechanistic studies revealed that ICG-001, overexpression of miR-96, or knockdown of EVI1 expression could restore the expression of miR-449a. The suppressive effect of miR-449a on the cell migration and tumor sphere formation was confirmed in NPC cells. Taken together, the miR-96/EVI1/miR-449a axis is a novel pathway involved in ICG-001-mediated inhibition of NPC cell migration and growth of the tumor spheres.

Introduction Traumatic brain injury (TBI) causes enormous morbidity and mortality in both adults and children. Children who survive TBI may have significant disability. It is not known if patterns of outpatient claim use among TBI survivors can be used to estimate patient functional status. Objectives and Approach The objectives of this study are to 1) link regional data sources to create a dataset containing information about the entire continuum of care for children with TBI and 2) identify patterns of outpatient medical needs that can serve as proxies for functional status. We probabilistically linked trauma registry and electronic medical record (EMR) data for children hospitalized after acute TBI at the two pediatric trauma centers in Colorado, Children’s Hospital Colorado (CHCO) and Denver Health (DH), with claims data from the Colorado All Payer Claims Database (APCD) (2009-2014). Probabilistic linkage was performed using LinkSolv (Strategic Matching, Inc., Morrisonville, NY). Results Of 737 hospitalized children, 387 (53%) linked to records in the APCD. Among the children in the linked cohort (213 at CHCO and 174 at DH), the median age was 5 years (interquartile range [IQR] 2 to 13) and 264 (68%) were male. Most had either mild (Glasgow Coma Scale [GCS] 13-15, 51%) or severe (GCS

Introduction Bloom Filters (BFs) are a scalable solution for probabilistic privacy-preserving record linkage but BFs can be compromised. Yao’s garbled circuits (GCs) can perform secure multi-party computation to compute the similarity of two BFs without a trusted third party. The major drawback of using BFs and GCs together is poor efficiency. Objectives and Approach We evaluated the feasibility of BFs+GCs using high capacity compute engines and implementing a novel parallel processing framework in Google Cloud Compute Engines (GCCE). In the Yao’s two-party secure computation protocol, one party serves as the generator and the other party serves as the evaluator. To link data in parallel, records from both parties are divided into chunks. Linkage between every two chunks in the same block is processed by a thread. The number of threads for linkage depends on available computing resources. We tested the parallelized process in various scenarios with variations in hardware and software configurations. Results Two synthetic datasets with 10K records were linked using BFs+GCs on 12 different software and hardware configurations which varied by: number of CPU cores (4 to 32), memory size (15GB – 28.8GB), number of threads (6-41), and chunk size (50-200 records). The minimum configuration (4 cores; 15GB memory) took 8,062.4s to complete whereas the maximum configuration (32 cores; 28.8GB memory) took 1,454.1s. Increasing the number of threads or changing the chunk size without providing more CPU cores and memory did not improve the efficiency. Efficiency is improved on average by 39.81% when the number of cores and memory on the both sides are doubled. The CPU utilization is maximized (near 100% on both sides) when the computing power of the generator is double the evaluator. Conclusion/Implications The PPRL runtime of BFs+GCs was greatly improved using parallel processing in a cloud-based infrastructure. A cluster of GCCEs could be leveraged to reduce the runtime of data linkage operations even further. Scalable cloud-based infrastructures can overcome the trade-off between security and efficiency, allowing computationally complex methods to be implemented.

Over 400 million years ago, the evolution of vertebrates gave rise to a life cycle in which the organism began to live longer particularly as an adult. To accommodate such a longer lifespan, the organism underwent adaptation, developing a mechanism for long-lived cellular homeostasis. This adaptation required a population of long-lived relatively quiescent somatic stem cells (SSCs) along with a more proliferative differentiated daughter cell population, and was necessary to safeguard the genetic attributes with which SSCs were endowed. Intriguingly, cAMP response element binding protein (CREB)-binding protein (CBP) and E1A-binding protein, 300 kDa (p300), the highly homologous Kat3 coactivators had diverged, through duplication of ancestral Kat3, immediately preceding the evolution of vertebrates, given that both CBP and p300 have been detected in nearly all vertebrates versus non-vertebrates. We now demonstrate that a relatively small, highly evolutionarily conserved, amino terminal 9 amino acid deletion in CBP versus p300, plays a critical role in allowing for both robust maintenance of genomic integrity in stem cells and the initiation of a feed-forward differentiation mechanism by tightly controlling the interaction of the nuclear receptor family with the Wnt signaling cascade in either an antagonistic or synergistic manner.

A Declaração dos BRICS na Cidade do Cabo em 10 de fevereiro de 2014 comprometeu os cinco signatários para um programa de cooperação em ciência, tecnologia e inovação (CTI). Foi tomada a decisão específica de alocar responsabilidades setoriais para cada parte: mudança climática e atenuação de catástrofes (Brasil); recursos hídricos e tratamento de poluição (Rússia); tecnologia geoespacial e suas aplicações (Índia); energias novas e renováveis, e eficiência energética (China); e astronomia (África do Sul). A ciência possui uma aparência de neutralidade, e como tal é uma importante aliada da diplomacia "soft", assim como o tênis de mesa na reaproximação entre Nixon e Mao em 1975. O mesmo não é exatamente verdade com respeito à tecnologia, com sua óbvia importância militar e com a inovação, que está intimamente conectada à competição econômica. A Declaração levanta, portanto, algumas questões interessantes. Qual é o presente estado da cooperação em CTI entre o grupo dos BRICS? Qual é a justificativa por trás da escolha dessas cinco áreas? Como essas áreas se alinham com a estratégia doméstica de cada país para CTI? Quais acordos bilaterais de CTI já estão em andamento, e o que eles têm produzido como resultados? Como a Declaração da Cidade do Cabo se alinha ou se compromete com o processo entre Índia, Brasil e África do Sul (IBSA)? As crescentes tensões geopolíticas (Mar Negro, Mar da China) irão limitar o escopo da cooperação em CTI? A análise de documentos e a bibliometria proporcionam algumas respostas às questões acima, porém o mais provável é que o fator decisivo sejam as dinâmicas geopolíticas globais.