Lower Survival Research Articles

Poly(ADP-Ribose) Polymerase (PARP) Inhibitors for Cancer Therapy: Advances, Challenges, and Future Directions.

Poly(ADP-ribose) polymerases (PARPs) are crucial nuclear proteins that play important roles in various cellular processes, including DNA repair, gene transcription, and cell death. Among the 17 identified PARP family members, PARP1 is the most abundant enzyme, with approximately 1-2 million molecules per cell, acting primarily as a DNA damage sensor. It has become a promising biological target for anticancer drug studies. Enhanced PARP expression is present in several types of tumors, such as melanomas, lung cancers, and breast tumors, correlating with low survival outcomes and resistance to treatment. PARP inhibitors, especially newly developed third-generation inhibitors currently undergoing Phase II clinical trials, have shown efficacy as anticancer agents both as single drugs and as sensitizers for chemo- and radiotherapy. This review explores the properties, characteristics, and challenges of PARP inhibitors, discussing their development from first-generation to third-generation compounds, more sustainable synthesis methods for discovery of new anti-cancer agents, their mechanisms of therapeutic action, and their potential for targeting additional biological targets beyond the catalytic active site of PARP proteins. Perspectives on green chemistry methods in the synthesis of new anticancer agents are also discussed.

Low magnesium levels and prognosis in newly diagnosed diffuse large B-cell lymphoma.

Magnesium (Mg) is an essential element involved in cellular metabolism. We demonstrated that in patients with diffuse large B-cell lymphoma (DLBCL) undergoing autologous stem cell transplant (SCT), those with a serum Mg < 2.0mg/dL at the time of transplant had worse outcomes. In this study, we aimed to learn the prognostic value of low serum Mg in patients with untreated DLBCL. We analyzed serum from 408 patients and tested 2 Mg cutpoints-low (<1.7mg/dL) and low normal (<2.0mg/dL), a range we found associated with lower survival in the SCT group. We found 3% of patients with low levels and 23% with low normal levels. Low normal serum Mg levels were associated with a higher stage at diagnosis, more extranodal involvement, higher international prognostic index score, lower overall survival (OS), and event-free survival. These data warrant testing Mg replacement to a target of >2.0mg/dL to learn if survival can be improved.

Local excision versus total mesorectal excision for rectal cancer patients with clinical complete or near-complete response after neoadjuvant chemoradiotherapy

PurposeLocal excision is an effective approach for managing rectal cancer exhibiting substantial regression after neoadjuvant chemoradiotherapy. The purpose of this study is to compare the outcomes between local excision and total mesorectal excision in rectal cancer patients achieving clinical complete or near-complete response after neoadjuvant chemoradiotherapy.MethodsThis is a retrospective cohort study that includes a consecutive series of rectal cancer patients who responded well to neoadjuvant chemoradiotherapy followed by surgery. A total of 180 rectal cancer patients at a single institution during a 12-year period are included. The main outcomes include short-term outcomes, oncological outcomes, and functional outcomes between the two groups.ResultsA total of 180 patients were included in the study. Sixty-one (33.9%) received local excision and 119 (66.1%) received total mesorectal excision. The baseline characteristics were generally balanced between the two groups. The local excision group demonstrated a significantly shorter operative time, less blood loss, and shorter hospital stay (p < 0.001). 3-year overall survival rates were 97.5% (95% CI, 0.93–1.00) and 95.5% (95% CI, 0.91–1.00) between the two groups (p = 0.38). The local excision group exhibited significantly higher 3-year local recurrence rates 15.7% (95% CI, 0.74–0.97) vs 4.2% (95% CI, 0.92–1.00) (p = 0.017), yet lower 3-year distant metastasis rates 9.6% (95% CI, 0.82–1.00) vs 12.6% (95% CI, 0.81–0.94) (p = 0.33) and lower 3-year disease-free survival rates 76.8% (95% CI, 0.64–0.92) vs 84.7% (95% CI, 0.78–0.92) (p = 0.56) comparing with the total mesorectal excision group. The local excision group demonstrated significantly better functional outcomes compared with the total mesorectal excision group (p < 0.001).ConclusionPatients who achieve either clinical complete or near-complete response after neoadjuvant chemoradiotherapy are suitable candidates for local excision. The local excision group demonstrated superior short-term and functional outcomes, and the oncological outcomes were not compromised.

Prognostic factors for premature cardiovascular disease mortality in Malaysia: a modelling approach using semi-parametric and parametric survival analysis with national health and morbidity survey linked mortality data.

Cardiovascular disease (CVD) is the leading cause of premature mortality worldwide. Despite existing research on CVD risk factors, the study of premature CVD mortality in Malaysia remains limited. This study employs survival analysis to model modifiable risk factors associated with premature CVD mortality among Malaysian adults. We utilised data from Malaysia's National Health and Morbidity Survey (NHMS) conducted in 2006, 2011, and 2015, linked with mortality records. The cohort comprised individuals aged 18 to 70 during the NHMS interview. Follow-up extended to 2021, focusing on CVD-related premature mortality between ages 30 and 70. We employed six survival models: a semi-parametric Cox proportional hazard (PH) and five parametric survival models, which were Exponential, Weibull, Gompertz, log-normal and log-logistic distributions using R software. The age standardized incidence rate (ASIR) of premature CVD mortality was calculated per 1000 person-years. Among 63,722 participants, 886 (1.4%) experienced premature CVD mortality, with an ASIR of 1.80 per 1000 person-years. The best-fit models (based on AIC value) were the stratified Cox model by age (semi-parametric) and the log-normal accelerated failure time (AFT) model (parametric). Males had higher risk (Hazard Ratio, HR = 2.68) and experienced 49% shorter survival time (Event Time Ratio, ETR = 0.51) compared to females. Compared to Chinese ethnicity, Indians, Malays, and other Bumiputera had higher HR and lower survival times. Rural residents and those with lower education also faced increased HRs and reduced survival times. Diabetes (diagnosed: HR = 3.26, ETR = 0.37; undiagnosed: HR = 1.63, ETR = 0.63), hypertension (diagnosed: HR = 1.84, ETR = 0.53; undiagnosed: HR = 1.46, ETR = 0.68), and undiagnosed hypercholesterolemia (HR = 1.31, ETR = 0.80) increased risk and decreased survival times. Additionally, current smoking and abdominal obesity elevated risk (HR = 1.38, 1.60) and shortened survival (ETR = 0.81, 0.71). The semi-parametric and parametric survival models both highlight the considerable impact of socioeconomic status and modifiable risk factors on premature CVD mortality, underscoring the imperative for targeted interventions to effectively mitigate these effects.

Serum Glial Fibrillary Acidic Protein Can Predict Cross-Sectional Vasculitis Activity by Reflecting Renal Involvement in Patients with Antineutrophil Cytoplasmic Antibody-Associated Vasculitis.

Background and Objectives: Glial fibrillary acidic protein (GFAP) is a type III intermediate filament protein primarily produced by cells in the central nervous system (CNS) and other major organs such as the kidneys. This study investigated whether serum GFAP could be used to estimate cross-sectional vasculitis activity presented via the Birmingham vasculitis activity score (BVAS) in patients with antineutrophil cytoplasmic antibody-associated vasculitis (AAV). Materials and Methods: This study included 74 patients with AAV. Clinical and laboratory data at diagnosis including BVAS and C-reactive protein (CRP) were reviewed. During follow-up, all-cause mortality and end-stage kidney disease (ESKD) were considered poor outcomes. Serum GFAP was measured from sera collected and stored at diagnosis. Results: The median age of the 74 patients was 63.5 years. Serum GFAP was inversely correlated with the cross-sectional BVAS (r = -0.373) and CRP (r = -0.320). It was also significantly correlated with general (r = -0.237) and renal (r = -0.335) manifestations among BVAS systemic items, and furthermore, among minor items of renal manifestation, correlating with sum scores for proteinuria (r = -0.409) and haematuria (r = -0.305). Additionally, compared with patients with serum GFAP > 194.9 pg/mL, those with serum GFAP ≤ 194.9 pg/mL showed a higher risk for progression to ESKD (relative risk 3.150) and a significantly lower cumulative ESKD-free survival rate. Conclusions: This study demonstrated the clinical potential of serum GFAP at diagnosis for predicting not only cross-sectional vasculitis activity through the anticipation of the extent of renal involvement but also future progression to ESKD in patients with AAV.

Assessing radioiodine therapy long-term outcomes in differentiated thyroid cancer using nomograms.

This study explored the role of radioiodine therapy (RAI) in low-risk thyroid cancer patients and examined the disease-specific survival (DSS) rates in a large cohort of differentiated thyroid cancer patients (DTC). We obtained patient data from SEER database. Patients who underwent total thyroidectomy were included and categorized into three groups based on histology: classical papillary thyroid carcinoma (C-PTC), follicular type variant carcinoma (FV-PTC), and follicular thyroid cancer (FTC). Patients with distant metastasis, tumor size ≥ 200mm, chemotherapy, or any type of radiation other than RAI were also excluded. A nomogram was developed and tested for discrimination and calibration. In total, 96,532 thyroid cancer cases were examined, including 59,460 C-PTC, 31,583 FV-PTC, and 5,489 FTC cases. Age > 65years and male sex were correlated with lower survival rates across the subtypes. In addition, extrathyroidal extension had a worse survival effect in patients with FTC. DSS rates were compared between patients who received RAI and those who did not, with a 3% difference in C-PTC (94% vs. 91%, p < 0.001), 2% in FV-PTC (92% vs. 90%, p < 0.001), and 1% in FTC (89% vs. 88%, p = 0.084) at 15years. The nomograms for long-term DSS showed high discriminatory abilities with C-indices of 0.815, 0.805, and 0.781 for C-PTC, FV-PTC, and FTC, respectively. The developed nomogram can be used in the treatment plan for patients with DTC. Our study emphasizes the prognostic factors for DTC and highlights the need for personalized treatment plans based on individual risk profiles.

European eel (Anguilla anguilla) survival modeling based on a 22-year capture-mark-recapture survey of a Mediterranean subpopulation.

Since the 1980s, the European eel (Anguilla anguilla) has declined by over 90% in recruitment across its European and North African distribution area. This diadromous fish spawns at sea and migrates into continental waters, where it grows for three to more than 30 years, depending on habitat conditions and location. During their growth, different habitat use tactics can locally influence the life-history traits of eels, including their survival rates. Thus, the spatio-temporal dimension of this species is crucial for management. Based on a rare Mediterranean long-term survey of more than 20 years (2001-2022) in an artificial drainage canal connected to a vast brackish lagoon (the Vaccarès lagoon), we aimed to study the dynamics of one subpopulation's life-history traits. We used Bayesian multistate capture-mark-recapture (CMR) models to assess the temporal variability in survival and abundance at both seasonal and inter-annual scales, considering life-stage structure. High survival rates and low detection probabilities were found for the undifferentiated and female yellow stages. In contrast, female silver eels exhibited lower survival rates and higher capture probabilities. Estimating detection probabilities and survival rates enabled accurate assessment of relative abundance across different life stages and time periods. Our findings indicated a substantial decrease in the abundance of undifferentiated and female yellow eels in the early 2000s, whereas the abundance of female silver eels remained consistently low yet stable throughout the study period. Considering the life stage seemed essential to study the dynamics of the eel during its continental growing period. The present results will provide key elements to propose and implement suitable sustainable environmental management strategies for eel conservation.

Record of albinism in European ground squirrel (Spermophilus citellus) (Rodentia: Sciuridae) in Slovakia

The European ground squirrel is a mammalian species whose populations are declining and has therefore become the target of various conservation projects in recent years. Currently, there are only a few dozen isolated populations in the Central European region. Increased interest in these animals has contributed to the first record of albino specimens of this species. A total of four juvenile individuals from two litters from one colony were observed, and in addition to the documentation itself, elementary behaviour notes were also made. All specimens did not survive to adulthood, confirming the low survival rate of aberrant individuals in the wild.

Androgen Deficiency, Associations and Survival of Men With Stage 4 and 5 Chronic Kidney Disease: A Cohort Study.

Anaemia is a key cause of morbidity in chronic kidney disease (CKD). Androgen deficiency (AD) in males can contribute to anaemia of all causes, including in CKD. We sought to examine the prevalence of AD in men with CKD, the extent to which it contributed to anaemia and whether it was independently associated with long-term survival. This cross-sectional observational study was conducted among males aged 18 years and over with CKD stages 4 and 5. The study analysed morning blood samples with regard to their full blood count, urea and electrolytes, albumin, lipids, testosterone (T) and sex hormone binding globulin, with calculation of free testosterone by mass action equation. Mortality data were obtained 15 years later for survival analysis. Among 322 patients with a mean age of 63 years, the overall prevalence of AD was 68.9%. There was a statistically significant negative correlation between erythropoiesis stimulating agent (ESA) dose and testosterone concentrations (Pearson correlation -0.193, p = 0.05). There was a positive correlation between haemoglobin (Hb) and free testosterone level among patients not on ESA therapy (Pearson correlation 0.331, p < 0.001). Kaplan-Meier plots showed p < 0.001 on log-rank analysis, indicating that AD was significantly associated with worse survival. However, in Cox regression analysis, free testosterone was not associated with survival (95% CI for free testosterone 0.997-1.000). AD is highly prevalent among this population, and increases further with older age and more severe CKD warranting haemodialysis. Association of lower Hb and higher ESA dose with lower T concentration might be causative, which has important pharmaco-economic as well as clinical implications. Lower survival in men with low T, more likely reflects overall poor health rather than causation. A properly constituted randomised controlled study evaluating the effect of native T replacement is warranted in men with CKD and AD.

Ecotoxicological and metabolomic investigation of chronic exposure of Daphnia magna (Straus, 1820) to yttrium environmental concentrations

In order to estimate the effects on aquatic organisms of long-term exposure to low doses of yttrium (Y) as a potential emerging contaminant, ecotoxicological and metabolomic data were collected on the model organism Daphnia magna, a keystone species in freshwater ecosystems. Following an initial acute toxicity assessment, a 21-day chronic exposure experiment was conducted using a sublethal concentration of 27 μg L⁻¹ of Y, corresponding to the effective concentrations inducing 10 % effect (EC10) value for mortality endpoint and simulating the environmental Y level in aquatic systems. Results from the 21-day two-factor experiment combining microcrustacean survival, growth and reproduction bioassays and targeted gas chromatography-mass spectrometry (GC–MS) metabolomics indicated significant adverse effects of chronic exposure to Y on D. magna. Daphnids exposed to Y exhibited a significantly lower survival at day 21, delayed the maturity stage, including their first breeding, and decreased clutch size. On the side of metabolomics, a clear and general increase over time of both the number and the level of detected metabolites in the hydroalcoholic extracts of the whole organisms was observed. However, emerging from this broad temporal pattern, several bioactive metabolites were identified (e.g., 2,4-di‑tert‑butylphenol, itaconic acid, 3-hydroxybutyric acid, and trehalose) whose levels in extracts are linked to the presence of Y. These results emphasize the necessity of considering low-dose, long-term exposure scenarios in environmental risk assessments of rare earth elements (REEs), which have often been overlooked in favour of higher concentration studies.

The Composition and Function of Intestinal Microbiota Were Altered in Farmed Bullfrog Tadpoles (Aquarana catesbeiana) during Metamorphosis

The bullfrog Aquarana catesbeiana is one of the main farmed frog species in China, with a low overall survival of farmed bullfrogs from hatching to harvest since bullfrog tadpoles are fragile during the metamorphosis period. The intestinal bacterial community can play crucial roles in animal development; however, little is known about the alteration of the gut microbial community of A. catesbeiana during metamorphosis. The present study used 16S rRNA amplicon sequencing to investigate the intestinal bacterial community in A. catesbeiana at four distinct developmental stages. Moreover, we determined the bullfrog’s body morphological parameters and the intestine histology at different developmental stages. The results showed a reduction in the total length and snout–vent length of A. catesbeiana during metamorphosis. The intestinal microbial composition of A. catesbeiana exhibited variation throughout the process of metamorphosis. The terrestrial stage showed shifts in the bacterial composition compared to the aquatic stages, including a reduction in Bacteroidetes and an increase in Firmicutes. Furthermore, the presence of Prevotella, Bifidobacterium, Leucobacter, Corynebacterium, Bulleidia, Dorea, Robinsoniella, and Clostridium in A. catesbeiana metamorphosis appears to be mainly related to the host’s epithelial cells’ height and total body mass. The results indicated that the intestinal microbial composition changed with the bullfrog–tadpole metamorphosis. The genera of Prevotella, Bifidobacterium, Leucobacter, Corynebacterium, Bulleidia, Dorea, Robinsoniella, and Clostridium might be potential probiotics.

7272 Antineoplastic Effect of Cabozantinib in Primary and Continuous Human Anaplastic Thyroid Cancer Cells

Abstract Disclosure: G. Elia: None. S. Ferrari: None. F. Ragusa: None. V. Mazzi: None. A. Patrizio: None. E. Balestri: None. C. Botrini: None. L. Rugani: None. E. Barozzi: None. C. La Motta: None. P. Fallahi: None. A. Antonelli: None. Anaplastic thyroid cancer (ATC) is a highly aggressive cancer with a low median survival of about 6-10 months. The standard treatment include surgery, external hyperfractionated radiation therapy, and chemotherapy. However, these therapies are not effective and new therapeutical approaches are needed. Tyrosine kinase inhibitors (TKI) are a class of drugs able to inhibit different pathways that results inappropriately activated in cancer cells.To date no studies evaluated the effect of cabozantinib in primary human ATC cells, and in ATC continuous cell line in vitro. We aimed to test the efficacy of cabozantinib a multiTKI acting against VEGFR 1,2, c-MET, and RET (recently approved for aggressive differentiated thyroid cancer) in primary human ATC cells, and in ATC continuous cell line (8305C, CAL-62) in vitro. Therefore, for primary ATC cells we collected surgical thyroidal tissues from five patients with ATC, from thyroid biopsy at the moment of first surgical operation, and we tested the effect of cabozantinib. Cabozantinib showed its efficacy in inhibiting cell proliferation, and also in increasing apoptosis (p &amp;lt; 0.05) both in primary and in continuous ATC cells. In conclusion, cabozantinib demonstrated a good antineoplastic activity in human ATC cells in vitro. These results open the way for a possible clinical use of cabozantinib in patients with ATC, and are important for a personalized therapeutic approach to be tailored to each patient avoiding the administration of ineffective drugs. Presentation: 6/1/2024

ABCG Transporters in the Adaptation of Rice to Salt Stresses

The ATP-binding cassette (ABC) proteins are a diverse family of transmembrane transporter proteins widely identified in various organisms. The ABCG transporters belong to the G subfamily of the ABC transporter family. Rarely research on ABCG transporters involved in salt tolerance of rice was found. In this study, the evolutionary relationships, conserved motifs, intra- and inter-species homologous genes, and cis-acting elements of ABCG subfamily members were analyzed, and the expression changes of these genes under salt stress at 0 h, 3 h, and 24 h were detected. Based on these results, the candidate gene OsABCG7, which is induced by salt stress, was selected for further studies. Yeast experiments confirmed that the OsABCG7 gene might be involved in the regulation of salt tolerance. The abcg7 mutant showed a higher degree of leaf wilting and a lower survival rate, exhibiting a salt-sensitive phenotype. Systematic analysis of this family in rice helps design effective functional analysis strategies and provides data support for understanding the role of ABCG transporters under salt stress.

Latent space representation of electronic health records for clustering dialysis-associated kidney failure subtypes

ObjectiveKidney failure manifests in various forms, from sudden occurrences such as Acute Kidney Injury (AKI) to progressive like Chronic Kidney Disease (CKD). Given its intricate nature, marked by overlapping comorbidities and clinical similarities—including treatment modalities like dialysis—we sought to design and validate an end-to-end framework for clustering kidney failure subtypes. Materials and methodsOur emphasis was on dialysis, utilizing a comprehensive dataset from the UK Biobank (UKB). We transformed raw Electronic Health Record (EHR) data into standardized matrices that incorporate patient demographics, clinical visit data, and the innovative feature of visit time-gaps. This matrix structure was achieved using a unique data cutting method. Latent space transformation was facilitated using a convolution autoencoder (ConvAE) model, which was then subjected to clustering using Principal Component Analysis (PCA) and K-means algorithms. ResultsOur transformation model effectively reduced data dimensionality, thereby accelerating computational processes. The derived latent space demonstrated remarkable clustering capacities. Through cluster analysis, two distinct groups were identified: CKD-majority (cluster 1) and a mixed group of non-CKD and some CKD subtypes (cluster 0). Cluster 1 exhibited notably low survival probability, suggesting it predominantly represented severe CKD. In contrast, cluster 0, with substantially higher survival probability, likely to include milder CKD forms and severe AKI. Our end-to-end framework effectively differentiates kidney failure subtypes using the UKB dataset, offering potential for nuanced therapeutic interventions. ConclusionsThis innovative approach integrates diverse data sources, providing a holistic understanding of kidney failure, which is imperative for patient management and targeted therapeutic interventions.

Liver Dysfunction and Systemic Inflammation Drive Organ Failures in Acute Decompensation of Cirrhosis: A Multicentric Study.

Hospitalized patients with acute decompensation (AD) of cirrhosis are at risk of progressing to acute-on-chronic liver failure (ACLF), significantly increasing their mortality. The aim of this study was to identify key predictors and patient trajectories predisposing to ACLF. In this multicenter, prospective study spanning 2 years, clinical, biochemical, and 90-day survival data were collected from 625 patients with AD (European Association for the Study of the Liver criteria) across North, South, and East India. We divided the cohort into a Derivation cohort (DC: 318 patients) and a Validation cohort (VC: 307 patients). Predictive models for pre-ACLF were derived, validated, and compared with established scores such as model for end-stage liver disease (MELD) 3.0 and chronic liver failure Consortium acute decompensation. Of 625 patients (mean age 49 years, 83% male, 77.5% with alcohol-related liver disease), 32.2% progressed to ACLF. Patients progressing to ACLF showed significantly higher bilirubin (10.9 vs 8.1 mg/dL), leukocyte counts (9,400 vs 8,000 per mm 3 ), international normalized ratio (1.9 vs 1.8), and MELD 3.0 (28 vs 25) but lower sodium (131 vs 134 mEq/L) and survival (62% vs 86%) compared with those without progression ( P < 0.05) in the DC. Consistent results were noted with alcohol-associated hepatitis, infection and hepatic encephalopathy as additional risk factors in VC. Liver failure at presentation (odds ratio: 2.4 [in DC], 6.9 [in VC]) and the 7-day trajectories of bilirubin, international normalized ratio, and MELD 3.0 significantly predicted ACLF progression ( P < 0.001). A new pre-ACLF model showed superior predictive capability (area under the curve of 0.71 in DC and 0.82 in VC) compared with MELD 3.0 and chronic liver failure Consortium acute decompensation scores ( P < 0.05). Approximately one-third of AD patients in this Indian cohort rapidly progressed to ACLF, resulting in high mortality. Early identification of patients at risk can guide targeted interventions to prevent ACLF.

Care Patterns and Outcomes for Intrahepatic Cholangiocarcinoma by Rurality of Patient Residence in a Midwestern State.

Intrahepatic cholangiocarcinoma (ICC) is the second most common malignancy of the liver and has the worst prognosis of any tumor arising from the liver, with a 5-year survival as low as 10%. However, whether the rurality of a patient's residence impacts care received and survival has not been well studied. We aimed to assess differences in care patterns associated with the rurality of patient's residences and their impact on survival outcomes, hypothesizing that patients in rural areas would experience lower survival. Adult patients diagnosed with ICC between 2010 and 2020 were identified in the Iowa Cancer Registry. Chi-square tests were used to compare values categorical variables by rural/urban status. Cox proportional hazards regression was used to determine associations with cancer-specific mortality. Of 672 patients diagnosed with ICC during the study period, 53%, 27%, and 21% resided in metropolitan, micropolitan, and rural areas, respectively. There were no significant differences in age, sex, stage at diagnosis, the proportion receiving chemotherapy within 12 weeks of diagnosis, and undergoing surgery across all groups. Additionally, the proportion receiving definitive care at a National Cancer Institute (NCI) designated center was comparable across the three groups (37% metro vs. 43% micro vs. 35% rural). However, rural residents had the highest proportion of traveling ≥ 50 miles for definitive care (22% metro vs. 41% micro vs. 56% rural). In multivariable analysis of patients with Stage 1-3 disease, younger age, receipt of chemotherapy, surgery, and definitive care at an NCI center were independently associated with decreased mortality risk. However, rural residence was not significantly associated with survival (HR: 0.64 [95% CI: 0.38-1.06]). Similar to other complex cancer diagnoses, we found that definitive care at an NCI center was associated with decreased mortality risk for patients with ICC. Although rural residence was not independently associated with survival in this cohort, rural residents traveled significantly longer distances to access definitive care. This highlights a crucial need to improve access to specialized centers for complex cancer care.

Mitotic spindle positioning protein serves as prognostic biomarker in patients with colorectal cancer

Background Colorectal cancer (CRC) ranks among the most aggressive types of cancer globally. Currently, clinical tumor prognostic biomarkers still lack accuracy. Mitotic spindle positioning (MISP) protein connects microtubules to the actin cytoskeleton and adhesive plaques, playing a critical role in spindle positioning, orientation, and the process of cell division. MISP can regulate the malignant biological functions of pancreatic cancer and intrahepatic cholangiocarcinoma and it acts as biomarker for prognosis, but its role in CRC remains unclear. Methods This study has collected 37 CRC tissue samples and 37 corresponding adjacent nontumor tissue samples, and 57 additional CRC tissues samples. Clinical data were obtained from the patients with CRC. MISP mRNA and protein expression levels were analyzed in normal control and CRC tissues using the GEPIA and Human Protein Atlas website. MISP protein levels in the collected tissues were analyzed using immunohistochemistry. Results MISP mRNA and protein expression levels were significantly increased in CRC tissues compared to adjacent nontumor tissues. Higher MISP protein levels were associated with distant metastasis, recurrence, and lower survival rates. Kaplan–Meier analysis showed that high expression levels of MISP protein were associated with recurrence and death in CRC patients. In addition, a high expression level of MISP protein, lymph node metastasis, and distance metastasis were risk factors for recurrence and a poor prognosis in patients with CRC. Conclusion Elevated MISP protein correlated with tumor metastasis, recurrence, and lower survival rates in patients with CRC, and thus, MISP has the potential to become a prognostic marker for CRC.

AP-2α decreases TMZ resistance of recurrent GBM by downregulating MGMT expression and improving DNA damage

AimsThe incidence of recurrent gliomas is high, exerting low survival rates and poor prognoses. Transcription factor AP-2α has been reported to regulate the progression of primary glioblastoma (GBM). However, the function of AP-2α in recurrent gliomas is largely unclear. MethodsThe expression of AP-2α and O6-methylguanine DNA-methyltransferase (MGMT) was detected in recurrent glioma tissues and cell lines by Western blots, the regulation mechanisms between AP-2α/MGMT promoter and RA/AP-2α promoter were studied by luciferase reporter assays, EMSA, and chIP assays. The effects of AP-2α and TMZ/RA treatment on cell viability in vitro and in vivo were investigated by MTT assays, γH2AX staining, comet assays and intracranial injection. Key findingsAP-2α expression negatively correlates with the expression of MGMT in glioma samples. AP-2α could directly bind with the promoter of the MGMT gene, suppresses transcriptional levels of MGMT and downregulate MGMT expression in TMZ-resistant U87MG-R and T98G cells, but TMZ treatment decreases AP-2α expression and increases MGMT expression. The extended TMZ treatment and increased TMZ concentrations reversed these effects. Moreover, AP-2α overexpression combines with TMZ to decrease cell viability, concurrently with improved DNA damage marker γH2AX. Furthermore, retinoic acid (RA) activates RAR/RXR heterodimers, which bind to RA-responsive elements (RAREs) of the AP-2α promoter, and activates AP-2α expression in recurrent glioma cells. Finally, in intracranial relapsed glioma mouse model, both RA and TMZ could retard tumor development and prolong the mouse survival. SignificanceAP-2α activation by gene overexpression or RA treatment reveals the suppressive effects on glioma relapse, providing a novel therapeutic strategy against malignant refractory gliomas.

Multi-omics analysis unveils the predictive value of IGF2BP3/SPHK1 signaling in cancer stem cells for prognosis and immunotherapeutic response in muscle-invasive bladder cancer

BackgroundMuscle invasive bladder cancer (MIBC) is a life-threatening malignant tumor characterized by high metastasis rates, poor prognosis, and limited treatment options. Immune checkpoint inhibitors (ICIs) targeting PD-1 and PD-L1 represent an emerging treatment for MIBC immunotherapy. However, the characteristics of patients likely to benefit from immunotherapy remain unclear.MethodsWe performed single-cell mass cytometry (CyTOF) analysis of 179,483 single cells to characterize potential immunotherapy-related cancer stem cells (CSCs)-like populations in the tumor microenvironment of 38 MIBC tissues. The upregulated expression of IGF2BP3 in CD274 + ALDH + CSC-like cells, which was associated with poor clinical prognosis, was analyzed by bulk RNA-sequencing data from an in-house cohort. The functional role of IGF2BP3 was determined through cell proliferation, colony formation, cell apoptosis and sphere formation assays. The regulation of SPHK1 expression by IGF2BP3 was investigated using methylated RNA immunoprecipitation sequencing (MeRIP-seq) and bulk RNA-sequencing (bulk RNA-seq). We further utilized single-nucleus RNA sequencing (snRNA-seq) data from 67,988 cells of 25 MIBC tissues and single-cell RNA sequencing (scRNA-seq) data from MIBC patient-derived organoids to characterize the molecular features of bladder cancer cells co-expressing IGF2BP3 and SPHK1. Spatial transcriptomics (ST) and co-detection by indexing (CODEX) analysis were used to describe the spatial distribution and interactions of IGF2BP3 + SPHK1 + bladder cancer cells and immune cells.ResultsA subset of CD274 + ALDH + CSC-like cells was identified, associating with immunosuppression and low survival rates in MIBC patients. IGF2BP3, an m6A reader gene, was found to be upregulated in the CD274 + ALDH + CSC-like cell population and linked to poor clinical prognosis in MIBC. Knockout of IGF2BP3 dramatically promoted cell apoptosis and reduced cell proliferation in T24 cells. By integrating MeRIP-seq and bulk RNA-seq analyses, we identified SPHK1 served as a substrate for IGF2BP3 in an m6A-dependent manner. Further snRNA-seq, scRNA-seq, ST, and CODEX analysis revealed a closer topographical distance between IGF2BP3 + SPHK1 + bladder cancer cells and exhausted CD8 + T cells, providing one explanation for the superior response to immunotherapy in IGF2BP3 + SPHK1 + bladder cancer cells-enriched patients. Finally, an ICI-associated signature was developed based on the enriched genes of IGF2BP3 + SPHK1 + bladder cancer cells, and its potential ability to predict the response to immunotherapy was validated in two independent immunotherapy cohort.ConclusionsOur study highlighted the critical involvement of the IGF2BP3/SPHK1 signaling in maintaining the stemness of CSCs and promoting MIBC progression. Additionally, these findings suggested that the IGF2BP3/SPHK1 signaling might serve as a biomarker for prognosis and immunotherapy response in MIBC.

Development and In vitro and In vivo efficacy investigation of multifunctional, targeted, theranostic liposomes for imaging and photodynamic therapy of glioblastoma

The challenge of effectively diagnosing and treating glioblastoma, which has a high incidence and low survival rate, remains unresolved. Consequently, research efforts have increased to develop specific and effective diagnostic and therapeutic agents. In this study, N-acetyl glucosamine-modified, IR780-encapsulated, PEG-coated, nanosized, 68Ga-labeled, neutral, and positively charged theranostic liposomes were developed for GLUT1 targeting for PET/CT and NIR imaging and PDT/PTT of GBM. Characterization, in vitro release profile, stability, radiolabeling efficiency, and labeling stability were evaluated. In vitro cell uptake studies were conducted in RG2 and U87 GBM cell lines, NIR images were obtained in the RG2 and U87 cell lines, and fluorescence microscope images were obtained in the RG2 cell line. PDT/PTT activities were assessed after exposure to NIR light (808 nm, 0.8–1 W/cm2). Cytotoxicity was studied in RG2, U87, and L929 cell lines. In vivo studies were performed in GBM model nude mice. The liposomes had an optimum mean particle size (181–193 nm), high encapsulation efficiency (79–84 %), and zeta potential (−5 to −7 mV). The maximum 68Ga labeling efficiency was achieved with an incubation at 80 °C, pH: 3.5, and 5 min. Liposomes remained stable for 30 days at 4 °C. While the intracellular uptake of targeted formulations was higher than non-targeted ones, it was similar for positively charged and neutral formulations. In U87 and RG2 cell lines, cell viability remained above 50 % for liposomal formulations at a concentration of 1.5 μg/mL exposed to NIR light (808 nm, 0.8–1 W/cm2). The formulations showed significant PDT/PTT activity under NIR light and lower cytotoxicity in the L929 than in RG2 and U87 cell lines. In vivo bioluminescence images and biodistribution studies in U87 tumor-bearing nude mice revealed higher tumor uptake of targeted, IR780-encapsulated liposomal formulations in tumor tissue compared to non-targeted ones. Liposomal formulations showed higher tumor uptake than the free IR780 solution. The data suggest that targeted liposomes have potential as theranostic agents for PDT/PTT and multifunctional glioblastoma imaging.