CPSF3 inhibition blocks pancreatic cancer cell proliferation through disruption of core histone mRNA processing.

Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease with limited effective treatment options, potentiating the importance of uncovering novel drug targets. Here, we target cleavage and polyadenylation specificity factor 3 (CPSF3), the 3' endonuclease that catalyzes mRNA cleavage during polyadenylation and histone mRNA processing. We find that CPSF3 is highly expressed in PDAC and is associated with poor prognosis. CPSF3 knockdown blocks PDAC cell proliferation and colony formation in vitro and tumor growth in vivo. Chemical inhibition of CPSF3 by the small molecule JTE-607 also attenuates PDAC cell proliferation and colony formation, while it has no effect on cell proliferation of nontransformed immortalized control pancreatic cells. Mechanistically, JTE-607 induces transcriptional readthrough in replication-dependent histones, reduces core histone expression, destabilizes chromatin structure, and arrests cells in the S-phase of the cell cycle. Therefore, CPSF3 represents a potential therapeutic target for the treatment of PDAC.

Revisiting Traditional Chinese Medicine in Hong Kong during the Influenza Epidemics in the 1950s and 1960s.

This paper examines the supply and utilization of traditional Chinese medicine (TCM) in Hong Kong during the influenza epidemics of the 1950s and 1960s. Existing narratives of TCM in Hong Kong have predominantly framed with within the dichotomy of Western medicine "Xiyi" and Chinese medicine "Zhongyi," portraying TCM as marginalized and nearly wiped out by colonial power. Departing from this binary opposition, this study views TCM as an autonomous space that had never been subjugated by the colonial power which opted for minimal interventionist approach toward TCM. By adopting diachronic and synchronic perspectives on Hong Kong's unique environment shaped by its colonial history and the geopolitics of the Cold War in East Asia, particularly its relationships with "China," this research seeks to reassess the role and status of TCM in post-World War II Hong Kong. In Hong Kong, along with other countries in East Asia, traditional medicine has ceded its position as mainstream medicine to Western medicine. Faced with the crisis of "extinction," Chinese medical professionals, including medical practitioners and merchant groups, persistently sought solidarity and "self-renewal." In the 1950s and 1960s, the colonial authorities heavily relied on private entities, including charity hospitals and clinics; furthermore, there was a lack of provision of public healthcare and official prevention measures against the epidemic influenza. As such, it is not surprising that the Chinese utilized TCM, along with Western medicine, to contain the epidemics which brought about an explosive surge in the number of patients from novel influenza viruses. TCM was significantly consumed during these explosive outbreaks of influenza in 1957 and 1968. In making this argument, this paper firstly provides an overview of the associations of Chinese medical practitioners and merchants who were crucial to the development of TCM in Hong Kong. Secondly, it analyzes one level of active provision and consumption of Chinese medicine during the two flu epidemics, focusing on the medical practices of TCM practitioners in the 1957 epidemic. While recognizing the etiologic agent or agents of the disease as influenza viruses, the group of Chinese medical practitioners of the Chinese Medical Society in Hong Kong adopted the basic principles of traditional medicine regarding influenza, such as Shanghanlun and Wenbingxue, to distinguish the disease status among patients and prescribe medicine according to correct diagnoses, which were effective. Thirdly, this paper examines the level of folk culture among the people, who utilized famous prescriptions of Chinese herbal medicine and alimentotherapy, in addition to Chinese patent medicines imported from mainland China. In the context of regional commercial network, this section also demonstrates how Hong Kong served as a sole exporting port of medicinal materials (e.g., Chinese herbs) and Chinese patent medicines from the People's Republic of China to capitalist markets, including Hong Kong, under the socialist planned or controlled economy in the 1950s and 1960s. It was not only the efficacy of TCM in restoring immunity and alleviating symptoms of the human body, but also the voluntary efforts of these Chinese medical practitioners who sought to defend national medicine "Guoyi," positioning it as complementary and alternative medicine to scientific medicine. Additionally, merchants who imported and distributed Chinese medicinal materials and national "Guochan" Chinese patent medicine played a crucial role, as did the people who utilized Chinese medicine, all of which contributed to making TCM thrive in colonial Hong Kong.

Data on the evaluation of the relation between ß-arrestin 2 and YAP phosphorylation in patient-derived colon cancer organoids.

The data presented in this article is related to a rapid communication entitled "ß-arrestin 2 suppresses the activation of YAP by promoting LATS kinase activity". This article describes the correlation of ß-arrestin 2 and YAP phosphorylation in patient-derived organoid models. Here, we analyzed 45 colon cancer organoids (CCOs) selected in the related research article to investigate the role of ß-arrestin 2 in YAP phosphorylation. Hematoxylin and eosin (H&E) staining and immunohistochemistry data showed that the CCOs maintained tissue architecture and histological features of their original cancer tissues. Moreover, mutation data detected from RNA-seq (RNA-sequencing) analysis showed that these CCOs retained the genetic features of their original colon cancer tissues as well. We also confirmed at the protein level that organoids expressing ß-arrestin 2 showed high expression of phosphorylated YAP. These organoid model studies strongly support the related research article that ß-arrestin 2 suppresses the activation of YAP in colon cancer.

Activation of the HSP27-AKT axis contributes to gefitinib resistance in non-small cell lung cancer cells independent of EGFR mutations.

PURPOSE: Although epidermal growth factor receptor (EGFR)-activating mutations in non-small cell lung cancer (NSCLC) usually show sensitivity to first-generation EGFR-tyrosine kinase inhibitors (TKIs), most patients relapse because of drug resistance. Heat shock protein 27 (HSP27) has been reported to be involved in the resistance of EGFR-TKIs, although the underlying mechanism is unclear. Here, we explore the mechanisms of HSP27-mediated EGFR TKI resistance and propose novel therapeutic strategies. METHODS: To determine the mechanism of HSP27 associated gefitinib resistance, differences were assessed using gefitinib-sensitive and -resistant NSCLC cell lines. In vivo xenograft experiments were conducted to elucidate the combinatorial effects of J2, a small molecule HSP27 inhibitor, and gefitinib. Analyses of human NSCLC tissues and PDX tissues were also used for comparison of HSP27 and phosphorylated AKT expression. RESULTS: Large-scale cohort analysis of NSCLC cases revealed that HSP27 expression correlated well with the incidence of EGFR mutations and affected patient survival. Increased pAKT and HSP27 was observed in gefitinib-resistant cells compared with gefitinib-sensitive cells. Moreover, increased phosphorylation of HSP27 by gefitinib augmented its protein stability and potentiated its binding activity with pAKT, which resulted in increased gefitinib resistance. However, in gefitinib-sensitive cells, stronger binding activity between EGFR and HSP27 was observed. Moreover, these phenomena occurred regardless of EGFR mutation including secondary mutations, such as T790M. AKT knockdown switched HSP27-pAKT binding to HSP27-EGFR, which promoted gefitinib sensitivity in gefitinib-resistant cells. Functional inhibition of HSP27 yielded sensitization to gefitinib in gefitinib-resistant cells by inhibiting the interaction between HSP27 and pAKT. CONCLUSIONS: Our results indicate that combination of EGFR-TKIs with HSP27 inhibitors may represent a good strategy to overcome resistance to EGFR-TKIs, especially in cancers exhibiting AKT pathway activation.

Immuno-genomic classification of colorectal cancer organoids reveals cancer cells with intrinsic immunogenic properties associated with patient survival.

BACKGROUND: The intrinsic immuno-ge7nomic characteristics of colorectal cancer cells that affect tumor biology and shape the tumor immune microenvironment (TIM) are unclear. METHODS: We developed a patient-derived colorectal cancer organoid (CCO) model and performed pairwise analysis of 87 CCOs and their matched primary tumors. The TIM type of the primary tumor was classified as immuno-active, immuno-exhausted, or immuno-desert. RESULTS: The gene expression profiles, signaling pathways, major oncogenic mutations, and histology of the CCOs recapitulated those of the primary tumors, but not the TIM of primary tumors. Two distinct intrinsic molecular subgroups of highly proliferative and mesenchymal phenotypes with clinical significance were identified in CCOs with various cancer signaling pathways. CCOs showed variable expression of cancer-specific immune-related genes such as those encoding HLA-I and HLA-II, and molecules involved in immune checkpoint activation/inhibition. Among these genes, the expression of HLA-II in CCOs was associated with favorable patient survival. K-means clustering analysis based on HLA-II expression in CCOs revealed a subgroup of patients, in whom cancer cells exhibited Intrinsically Immunogenic Properties (Ca-IIP), and were characterized by high expression of signatures associated with HLA-I, HLA-II, antigen presentation, and immune stimulation. Patients with the Ca-IIP phenotype had an excellent prognosis, irrespective of age, disease stage, intrinsic molecular type, or TIM status. Ca-IIP was negatively correlated with intrinsic E2F/MYC signaling. Analysis of the correlation between CCO immuno-genotype and TIM phenotype revealed that the TIM phenotype was associated with microsatellite instability, Wnt/ß-catenin signaling, APC/KRAS mutations, and the unfolded protein response pathway linked to the FBXW7 mutation in cancer cells. However, Ca-IIP was not associated with the TIM phenotype. CONCLUSIONS: We identified a Ca-IIP phenotype from a large set of CCOs. Our findings may provide an unprecedented opportunity to develop new strategies for optimal patient stratification in this era of immunotherapy.

A one-stop microfluidic-based lung cancer organoid culture platform for testing drug sensitivity.

Microfluidic devices as translational research tools provide a potential alternative to animal experiments due to their ability to mimic physiological parameters. Several approaches that can be used to predict the efficacy or toxicity of anticancer drugs are available. In general, standard cell culture systems have the advantages of being relatively cost-effective, having high-throughput capability, and providing convenience. However, these models are inadequate to accurately recapitulate the complex organ-level physiological and pharmacological responses. Here, we present a one-stop microfluidic device enabling both 3-dimensional (3D) lung cancer organoid culturing and drug sensitivity tests directly on a microphysiological system (MPS). Our platform reproducibly yields 3D lung cancer organoids in a size-controllable manner and demonstrates for the first time the production of lung cancer organoids from patients with small-cell lung cancer. Lung cancer organoids derived from primary small-cell lung cancer tumors can rapidly proliferate and exhibit disease-specific characteristics in our MPS. Cisplatin and etoposide, the standard regimen for lung cancer, showed increased apoptosis induction in a concentration-dependent manner, but the organoids contained chemo-resistant cells in the core. We envision that this system may provide important information to guide therapeutic approaches at the preclinical level.

Patient-derived lung cancer organoids as in vitro cancer models for therapeutic screening.

Lung cancer shows substantial genetic and phenotypic heterogeneity across individuals, driving a need for personalised medicine. Here, we report lung cancer organoids and normal bronchial organoids established from patient tissues comprising five histological subtypes of lung cancer and non-neoplastic bronchial mucosa as in vitro models representing individual patient. The lung cancer organoids recapitulate the tissue architecture of the primary lung tumours and maintain the genomic alterations of the original tumours during long-term expansion in vitro. The normal bronchial organoids maintain cellular components of normal bronchial mucosa. Lung cancer organoids respond to drugs based on their genomic alterations: a BRCA2-mutant organoid to olaparib, an EGFR-mutant organoid to erlotinib, and an EGFR-mutant/MET-amplified organoid to crizotinib. Considering the short length of time from organoid establishment to drug testing, our newly developed model may prove useful for predicting patient-specific drug responses through in vitro patient-specific drug trials.

KIF3A binds to ß-arrestin for suppressing Wnt/ß-catenin signalling independently of primary cilia in lung cancer.

Aberrant Wnt/ß-catenin signalling is implicated in the progression of several human cancers, including non-small cell lung cancer (NSCLC). However, mutations in Wnt/ß-catenin pathway components are uncommon in NSCLC, and their epigenetic control remains unclear. Here, we show that KIF3A, a member of the kinesin-2 family, plays a role in suppressing Wnt/ß-catenin signalling in NSCLC cells. KIF3A knockdown increases both ß-catenin levels and transcriptional activity with concomitant promotion of malignant potential, such as increased proliferation and migration and upregulation of stemness markers. Because KIF3A binds ß-arrestin, KIF3A depletion allows ß-arrestin to form a complex with DVL2 and axin, stabilizing ß-catenin. Although primary cilia, whose biogenesis requires KIF3A, are thought to restrain the Wnt response, pharmacological inhibition of ciliogenesis failed to increase ß-catenin activity in NSCLC cells. A correlation between KIF3A loss and a poorer NSCLC prognosis as well as ß-catenin and cyclin D1 upregulation further suggests that KIF3A suppresses Wnt/ß-catenin signalling and tumourigenesis in NSCLC.