Min Liu

Background: Patients with anaplastic lymphoma kinase (ALK)-positive malignancies increasingly rely on ALK inhibitors (ALKis) as first-line therapy, yet thromboembolism (TE) across the ALKi family and its impact on patients’ safety remain incompletely characterized, particularly in the real-world populations. Objective: The aim of this study was to evaluate the real-world TE risks associated with ALKis using pharmacovigilance data mining and to identify and characterize the high-risk contributors to TE. Methods: We extracted 52,808 adverse event (AE) reports from the FDA Adverse Event Reporting System (2013–2024), including 17,124 ALKi-treated patients. TE signal strength was calculated via disproportionality analysis and presented by the lower limit of reporting odds ratio (ROR ₀₂₅ ) and information component (IC ₀₂₅ ). Multivariate logistic regression and time-to-onset (TTO) analyses were conducted to characterize the risk factors. Results: Among ALKi users, 251 TE cases were identified. Crizotinib exhibited significant TE safety signals for embolism (ROR ₀₂₅ = 2.66; IC0 ₂₅ = 1.19), pulmonary artery thrombosis (ROR ₀₂₅ = 4.94; IC0 ₂₅ = 1.01), pulmonary thrombosis (ROR ₀₂₅ = 1.29; IC0 ₂₅ = 0.13), and venous thrombosis limb (ROR ₀₂₅ = 1.66; IC0 ₂₅ = 0.08). Older age (OR = 1.006, P = 2.40e-06), male sex (OR = 1.47, P = 2.00e-16), and crizotinib use (OR = 2.79, P = 1.59e-05) independently predicted fatal TE outcomes. Female patients <65 years experienced earlier TE onset (median 14 vs. 44–53 days; P = 0.006). Conclusion: This study validates the elevated TE safety signal strength of crizotinib in the real-world settings, while no disproportionate TE signals were detected for other ALKi in the FAERS database. Clinicians could prioritize agent-specific risks, especially in older male patients and during early treatment phases.

Mpox (monkeypox) infection cases increased recently in non-Mpox outbreak areas, potentially causing an international threat. The desire to defend against a potential outbreak has led to renewed efforts to develop Mpox vaccines. In this report, mice were immunized with various doses of modified vaccinia virus Ankara (MVA) to evaluate the cross-reactive immune response of MVA immunization against protective antigens of the current monkeypox virus. We demonstrated that MVA induced specific antibodies against protective antigens (A29, A35, B6, M1, H3, and I1), mediating the neutralization abilities against the MVA and the monkeypox virus (MPXV). Moreover, recombinant protective antigens of the MPXV elicited cross-binding and cross-neutralizing activities for MVA. Hence, the MVA induced cross-reactive immune responses, which may guide future efforts to develop vaccines against the recent MPXV. Notably, compared to the other protective antigens, the predominant A29 and M1 antigens mediated higher cross-neutralizing immune responses against the MVA, which could serve as antigen targets for novel orthologous orthopoxvirus vaccine.

Glioblastoma is the most common central nervous system cancer and is responsible for approximately one-half of all malignant brain tumors. With a 5-year survival rate of 6.8% and a median observed survival time of only 8 months, recurrent glioblastoma is among the world’s most lethal malignancies. Temozolomide (TMZ) has become a standard chemotherapeutic for glioblastoma due to its alkylating effects and ability to cross the blood-brain barrier. Unfortunately, many patients with glioblastoma develop resistance to TMZ, severely limiting therapeutic options. Recent studies have found a link between ectopic connexin 43 (Cx43) and TMZ-resistant glioblastoma. Further studies have shown success in overcoming this resistance by combining TMZ with alpha carboxyl terminus 1 (αCT1) peptide, a clinically tested Cx43 carboxyl terminus mimetic peptide inhibitor; however, because of the short half-life of αCT1, therapeutic potential is limited, and new approaches are needed. In this study, we hypothesized that simultaneously targeting Cx43 with αCT1 and phosphoinositide 3-kinase (PI3K) using PI3K isoform-selective inhibitors would synergistically sensitize glioblastoma to TMZ treatment because p110β, one of the four PI3K catalytic subunits, is a prime survival factor for TMZ-resistant glioblastoma. We cultured newly dissected primary glioblastoma cells, glioblastoma cell lines, and glioblastoma stem cells and dosed them with various combinations of αCT1, TMZ, and one of two clinically tested p110β inhibitors (TGX-221 or GSK2636771). Cell viability was measured using an MTS viability assay. Glioblastoma cells that expressed high levels of Cx43 and p110β experienced a significant decrease in viability when dosed with a triple combination. The additive properties of these combinations were measured using an excess over Bliss (EOB) score derived from Bliss independence modeling, which showed synergistic effects as high as 81% for triple therapy combinations in TMZ-resistant cell lines. The EOB was significantly higher in triple therapies than the double or single therapies. To support our findings, we used the same combinations in primary glioblastoma cells and measured the activity of cleaved caspase 3/7 using a luminescence assay. There was a synergistic increase in caspase 3/7 activity in the triple therapy that was not seen in any of the other combinations. Identical treatment in non-pathological human astrocytes suggests that our drug combinations do not exacerbate non-selective TMZ toxicity in the brain. Collectively, our data demonstrate that simultaneous inhibition of Cx43 and PI3K is an effective approach for overcoming TMZ resistance in glioblastoma. Citation Format: Kasen R. Hutchings, Kevin J. Pridham, Min Liu, Joseph Owens, Zhi Sheng. Synergistic effects of connexin 43 inhibitor and PI3K isoform-selective inhibitors in temozolomide-resistant glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3243.

P-TEFb modulates RNA polymerase II elongation through alternative interaction with negative and positive regulation factors. While inactive P-TEFbs are mainly sequestered in the 7SK snRNP complex in a chromatin-free state, most of its active forms are in complex with its recruitment factors, Brd4 and SEC, in a chromatin-associated state. Thus, switching from inactive 7SK snRNP to active P-TEFb (Brd4/P-TEFb or SEC/P-TEFb) is essential for global gene expression. Although it has been shown that cellular signaling stimulates the disruption of 7SK snRNP, releasing dephosphorylated and catalytically inactive P-TEFb, little is known about how the inactive released P-TEFb is reactivated. Here, we show that the Cdk9/CycT1 heterodimer released from 7SK snRNP is completely dissociated into monomers in response to stress. Brd4 or SEC then recruits monomerized Cdk9 and CycT1 to reassemble the core P-TEFb. Meanwhile, the binding of monomeric dephosphorylated Cdk9 to either Brd4 or SEC induces the autophosphorylation of T186 of Cdk9. Finally, the same mechanism is employed during nocodazole released entry into early G1 phase of cell cycle. Therefore, our studies demonstrate a novel mechanism by which Cdk9 and CycT1 monomers are reassembled on chromatin to form active P-TEFb by its interaction with Brd4 or SEC to regulate transcription.

The spotted box crab Calappa philargius (Calappidae) is an increasingly consumed species in China, mainly sourced from a claw-only fishery. Being a not well-characterized species in the literature, this study is warranted to inform sustainable management approaches. Here we report on the first in-depth overview on biology and fishery dynamics of C. philargius in the southern Taiwan Strait of China. Whole body crabs (N = 1,009) were collected monthly from January to December 2019 from trawlers, operating in the southern Taiwan Strait; sample collection was absent from May to July because of the national fishing moratorium regulation. Sex ratio, growth pattern, size at maturity and fecundity were estimated. The overall male: female ratio was 1: 1.47, significantly differing from the 1: 1 (p < 0.01); monthly sex ratio variations were also significant (p < 0.01), suggesting possible seasonal sexual segregation. We found the carapace widths (CW) of males were significantly larger than those of females (p < 0.01). Based on the occurrence and percentages of berried females, we identified twin spawning events in January–April and August–October with the overall peak occurring in February. Size at 50% female maturity was 11.47 cm CW. Female absolute fecundity was significantly related to CW via a power function relationship (N = 14, p < 0.01). The estimated average claw yield was 36.28 ± 3.07% of the whole body weight (N = 95), irrespective of sex and size (p > 0.5). Capture and trade data of C. philargius using trawl and trap fishing gears were documented to characterize claw-only fishery dynamics for future research and sustainable use.

Capicua (Cic), a transcriptional repressor frequently mutated in brain cancer oligodendroglioma, is highly expressed in adult neurons. However, its function in the dendritic growth of neurons in the hippocampus remains poorly understood. Here, we confirmed that Cic was expressed in hippocampal neurons during the main period of dendritogenesis, suggesting that Cic has a function in dendrite growth. Loss-of-function and gain-of function assays indicated that Cic plays a central role in the inhibition of dendritic morphogenesis and dendritic spinesin vitro. Further studies showed that overexpression of Cic reduced the expression of Ets in HT22 cells, whilein vitroknockdown of Cic in hippocampal neurons significantly elevated the expression of Ets. These results suggest that Cic may negatively control dendrite growth through Ets, which was confirmed by ShRNA knockdown of either Etv4 or Etv5 abolishing the phenotype of Cic knockdown in cultured neurons. Taken together, our results suggest that Cic inhibits dendritic morphogenesis and the growth of dendritic spines through Ets.

The aetiology of inflammatory bowel disease (IBD) is a multifactorial interplay between heredity and environment^1,2. Here we report that deficiency in SETDB1, a histone methyltransferase that mediates the trimethylation of histone H3 at lysine 9, participates in the pathogenesis of IBD. We found that levels of SETDB1 are decreased in patients with IBD, and that mice with reduced SETDB1 in intestinal stem cells developed spontaneous terminal ileitis and colitis. SETDB1 safeguards genome stability³, and the loss of SETDB1 in intestinal stem cells released repression of endogenous retroviruses (retrovirus-like elements with long repeats that, in humans, comprise approximately 8% of the genome). Excessive viral mimicry generated by motivated endogenous retroviruses triggered Z-DNA-binding protein 1 (ZBP1)-dependent necroptosis, which irreversibly disrupted homeostasis of the epithelial barrier and promoted bowel inflammation. Genome instability, reactive endogenous retroviruses, upregulation of ZBP1 and necroptosis were all seen in patients with IBD. Pharmaceutical inhibition of RIP3 showed a curative effect in SETDB1-deficient mice, which suggests that targeting necroptosis of intestinal stem cells may represent an approach for the treatment of severe IBD.

Ubiquilin4 (Ubqln4), a member of the UbL-UBA protein family, serves as an adaptor in the degradation of specific substrates via the proteasomal pathway. However, the biological function of Ubqln4 remains largely unknown, especially in cancer. Here, we reported that Ubqln4 was downregulated in gastric cancer tissues and functioned as a tumor suppressor by inhibiting gastric cancer cell proliferation in vivo and in vitro. Overexpression of Ubqln4-induced cellular senescence and G1-S cell cycle arrest in gastric cancer cells and activated the p53/p21 axis. Moreover, Ubqln4 regulated p21 through both p53-dependent and p53-independent manners. Ubqln4 interacted with RNF114, an E3 ubiquitin ligase of p21, and negatively regulated its expression level, which in turn stabilized p21 by attenuating proteasomal degradation of p21. These effects of Ubqln4 were partly abrogated in gastric cancer cells upon silencing of p21. Our findings not only establish the anti-tumor potential of Ubqln4 in gastric cancer but also reveal a role for Ubqln4 in regulation of the cell cycle and cellular senescence via stabilizing p21.