Unfavorable Clinical Outcome Research Articles

ALKB homolog 5 (ALKBH5)-induced circPUM1 upregulation facilitated the progression of neuroblastoma via miR-423–5p/PA2G4 axis

BackgroundThe oncogenic role of circPUM1 has been revealed in multiple cancers. Nevertheless, the specific role and molecular mechanism of circPUM1 in neuroblastoma (NB) have never been reported. MethodsThe expression of genes was detected using RT-qPCR and Western Blot assay. The proliferation, migration, and invasion of NB cells were evaluated by CCK-8 and Transwell assays. Besides, mouse model was established to evaluate the effect of circPUM1 on the progression of NB. The interaction among genes was verified through RIP, MeRIP, or Luciferase reporter assay. ResultsThrough our investigation, it was discovered that circPUM1 expression was abnormally elevated in NB tissues and the abundance of circPUM1 was correlated with unfavorable clinical outcomes in NB patients. Besides, the viability and mobility of NB cells as well as NB tumor growth were suppressed by silencing circPUM1. Moreover, bioinformatics prediction and experimental verification demonstrated that circPUM1 was a sponge for miR-423–5p which further targeted proliferation-associated protein 2G4 (PA2G4). The oncogenic effect of circPUM1 on NB was exerted through suppressing miR-423–5p to elevate PA2G4 expression. Finally, we investigated the transcriptional factor causing the upregulation of circPUM1 in NB. The result was that ALKB homolog 5 (ALKBH5), an m6A demethylase, suppressed the m6A modification of circPUM1 and caused the elevation of circPUM1 expression in NB. ConclusionALKBH5 induced the upregulation of circPUM1 to accelerate the development of NB through regulating miR-423–5p/PA2G4 axis.

A Novel Gene List Identifies Tumors with a Stromal-Mesenchymal Phenotype and Worse Prognosis in Gastric Cancer.

Molecular biomarkers that predict disease progression can help identify tumor subtypes and shape treatment plans. In this study, we aimed to identify robust biomarkers of prognosis in gastric cancer based on transcriptomic data obtained from primary gastric tumors. Microarray, RNA sequencing, and single-cell RNA sequencing-based gene expression data from gastric tumors were obtained from public databases. Freshly frozen gastric tumors (n = 42) and matched FFPE (formalin-fixed, paraffin-embedded) (n = 40) tissues from a Turkish gastric cancer cohort were used for quantitative real-time PCR and immunohistochemistry-based assessments of gene expression, respectively. A novel list of 20 prognostic genes was identified and used for the classification of gastric tumors into two major tumor subgroups with differential stromal gene expression ("Stromal-UP" (SU) and "Stromal-DOWN" (SD)). The SU group had a more mesenchymal profile with an enrichment of extracellular matrix-related gene sets and a poor prognosis compared to the SD group. Expression of the genes within the signature correlated with the expression of mesenchymal markers ex vivo. A higher stromal content in FFPE tissues was associated with shorter overall survival. A stroma-rich, mesenchymal subgroup among gastric tumors identifies an unfavorable clinical outcome in all cohorts tested.

SAT-525 - Detection of candida species in hospitalized patients with decompensated liver cirrhosis and ascites indicates an unfavorable clinical course and outcome

SAT-525 - Detection of candida species in hospitalized patients with decompensated liver cirrhosis and ascites indicates an unfavorable clinical course and outcome

COVID-19 Vaccination and Clinical Outcomes at a Secondary Referral Hospital During the Delta Variant-dominant Period in West Sumatra, Indonesia.

The second wave of coronavirus disease 2019 (COVID-19) cases in Indonesia, during which the Delta variant predominated, took place after a vaccination program had been initiated in the country. This study was conducted to assess the impact of COVID-19 vaccination on unfavorable clinical outcomes including hospitalization, severe COVID-19, intensive care unit (ICU) admission, and death using a real-world model. This single-center retrospective cohort study involved patients with COVID-19 aged ≥18 years who presented to the COVID-19 emergency room at a secondary referral teaching hospital between June 1, 2021 and August 31, 2021. We used a binary logistic regression model to assess the effect of COVID-19 vaccination on unfavorable clinical outcomes, with age, sex, and comorbidities as confounding variables. A total of 716 patients were included, 32.1% of whom were vaccinated. The elderly participants (≥65 years) had the lowest vaccine coverage among age groups. Vaccination had an effectiveness of 50% (95% confidence interval [CI], 25 to 66) for preventing hospitalization, 97% (95% CI, 77 to 99) for preventing severe COVID-19, 95% (95% CI, 56 to 99) for preventing ICU admission, and 90% (95% CI, 22 to 99) for preventing death. Interestingly, patients with type 2 diabetes had a 2-fold to 4-fold elevated risk of unfavorable outcomes. Among adults, COVID-19 vaccination has a moderate preventive impact on hospitalization but a high preventive impact on severe COVID-19, ICU admission, and death. The authors suggest that relevant parties increase COVID-19 vaccination coverage, especially in the elderly population.

Biomarker and genomic analyses reveal molecular signatures of non-cardioembolic ischemic stroke

Acute ischemic stroke (AIS) is a major cause of disability and mortality worldwide. Non-cardioembolic ischemic stroke (NCIS), which constitutes the majority of AIS cases, is highly heterogeneous, thus requiring precision medicine treatments. This study aimed to investigate the molecular mechanisms underlying NCIS heterogeneity. We integrated data from the Third China National Stroke Registry, including clinical phenotypes, biomarkers, and whole-genome sequencing data for 7695 patients with NCIS. We identified 30 molecular clusters based on 63 biomarkers and explored the comprehensive landscape of biological heterogeneity and subpopulations in NCIS. Dimensionality reduction revealed fine-scale subpopulation structures associated with specific biomarkers. The subpopulations with biomarkers for inflammation, abnormal liver and kidney function, homocysteine metabolism, lipid metabolism, and gut microbiota metabolism were associated with a high risk of unfavorable clinical outcomes, including stroke recurrence, disability, and mortality. Several genes encoding potential drug targets were identified as putative causal genes that drive the clusters, such as CDK10, ERCC3, and CHEK2. We comprehensively characterized the genetic architecture of these subpopulations, identified their molecular signatures, and revealed the potential of the polybiomarkers and polygenic prediction for assessing clinical outcomes. Our study demonstrates the power of large-scale molecular biomarkers and genomics to understand the underlying biological mechanisms of and advance precision medicine for NCIS.

Prospective assessment of platelet function in patients undergoing elective resection of glioblastoma multiforme

This prospective study was aimed to test changes in hemostasis in patients with GBM, occurring at baseline (before surgery, time 0, T0) and 2 (T2), 24 (T24), and 48-hour (T48) after surgery. We enrolled consecutive patients subjected to GBM resection (GBR group; N = 60), laparoscopic colon cancer resection (comparative CCR group; N = 40), and healthy blood donors (HBD group; N = 40). We performed 1. conventional coagulation tests 2. ROTEM (rotational thromboelastometry) parameters and 3. platelet function tests, including PFA-200 closure time when stimulated by collagen/epinephrine (COL-EPI) and ROTEM platelet, using three different activators (arachnoid acid in ARATEM, adenosine diphosphate in ADPTEM, and thrombin receptor-activating peptide-6 in TRAPTEM). Variables associated with unfavorable 1-year clinical outcome were investigated, too. We observed in GBR patients that platelet aggregometry, as assessed by ROTEM platelet parameters, was significantly impaired along with a shortened closure time. These changes were evident from T0 to T48. A decreased area under the aggregation curve in TRAPTEM was associated with improved survival (adjusted odd ratio (95% CI), 1.03 (1.01–1.06)). This study suggests that patients with GBM presented a decreased platelet aggregation from before surgery and thorough the postoperative period. Decreased platelet aggregation improved clinical outcome.

Doppler-derived cardiac performance and evaluation of alternative measures of AV conduction to predict unvfavuorable haemodynamics in patients with long PR (DEEPER PR Study)

Abstract Funding Acknowledgements Type of funding sources: None. Background Prolonged atrio-ventricular (AV) conduction has been shown to compromise cardiac performance. Long PR relates to unfavorable clinical outcome and even increased mortality. The PR interval is traditionally used to describe AV conduction, but this parameter comprises inter-atrial and AV conduction delays and may therefore not optimally reflect hemodynamic consequences of prolonged AV conduction. Ventricular conduction delay also impacts ventricular filling and cardiac performance, while delayed inter-atrial activation may alleviate LV hemodynamic compromise. It is unknown, which parameter of AV conduction relates closest to cardiac performance. The objective of this study is to characterize segments of the PR interval in patients with long PR vs. matched controls with normal PR and to investigate the association of various measures of AV conduction to Doppler-derived cardiac performance. Methods 52 hospitalized pts with AVB I° (group A), and 52 controls (matched according to age, gender, LVEF, group B) were studied in detail by digitalized 12 lead ECG and 2-D, Doppler, and tissue Doppler echocardiography. Segments of PR were compared between matched groups. Seven measures of AV conduction were correlated to LV and RV filling fraction, total isovolumic times, and Tei indices and ranked by logistic regression and ROC-AUC in the combined patient cohort. Results PR was 227.5 (212;251)ms (A) vs. 182 (164;190)ms (B); p<0.001). P wave duration was 127(114;136)ms (A) vs. 113(102.5;126.8)ms (B); p<0.001 with longer RA and LA activation in group A. Reduced LV filling fraction (PQ Δ0.045, PePV6 Δ0.050, AUC 0.681 vs. 0.719), prolonged combined LV isovolumic times (PR: Δ19ms, 15.8%, PePV6: Δ31ms, 26.9%, AUC 0.619 vs 0.698), and higher LV Tei indices (PR: Δ0.045, 10.3%, PePV6: Δ0.083, 19.8%, AUC 0.566 vs 0.626) were best predicted by the variable PePV6 (interval between the end of P in V1 and Rpeak in V6) according to bivariate correlations, logistic regression analyses, and ROC-AUC. Applying PePV6 instead of PR re-categorized 20% of pts as having prolonged vs normal AV conduction. Prolonged PePV6 was also best suited to predict worse RV hemodynamics. Although significant linear correlations were demonstrated, a non-linear component of the association between AV timing and LV filling appeard to be present with pronounced impairment of cardiac performance beyond the 3rd quartile of PePV6 (148ms), respectively. Conclusion Prolonged PR comprises prolonged atrial activation (30%) and delayed AV conduction (70%). In this study, PePV6, a novel parameter eliminating P wave duration and accounting for initial LV activation, was best suited to identify compromise of Doppler-derived LV and RV hemodynamics by delayed AV conduction. PePV6 prolongation beyond the 3rd quartile may particularly affect cardiac performance. These findings inform future clinical studies and should give rise to investigation of PePV6 in larger patient populations.

Implications of the Mitral Leaflet Coaptation Pattern on Clinical Outcomes in Patients With Functional Mitral Regurgitation

The classification of secondary mitral regurgitation (MR) is based on atrial functional MR (AFMR) or ventricular functional MR (VFMR) and volume changes, but the mitral leaflet coaptation angle also contributes to the MR mechanism. The clinical implications of the coaptation angle on cardiovascular (CV) outcomes have not been well evaluated. A total of 469 consecutive patients (265 AFMR vs 204 VFMR) with more than moderate MR were evaluated for the occurrence of heart failure, mitral valve operations, and CV death. The coaptation angle was assessed by measuring the internal angle between both leaflets at mid-systole using the apical 3-chamber view. A coaptation angle ≥130° was classified as leaflet flattening, and an angle <130° was classified as leaflet tethering. AFMR and VFMR were associated with higher frequencies of leaflet flattening and tethering, respectively. AFMR was more likely to be associated with older age, atrial fibrillation, and preserved ejection fraction, all of which were related to leaflet flattening. During a follow-up of 2.3years, 83 patients had heart failure (17.7%), 21 patients underwent mitral valve operations (4.5%), and 34 patients died (7%). Compared with leaflet tethering, leaflet flattening was more significantly related to CV events, whereas CV event rates were less markedly different in A/VFMR. Irrespective of A/VFMR, leaflet flattening and atrial fibrillation were associated with a higher frequency of CV events. Adjusted analysis showed that leaflet flattening remained an independent predictor of CV events (hazard ratio 3.5, 95% confidence interval 1.11 to 4.88, p=0.003), whereas A/VFMR did not. In conclusion, the leaflet coaptation angle in patients with functional MR could provide risk stratification superior to that of A/VFMR. Leaflet flattening appears to be associated with unfavorable clinical outcomes.

Implications of CRNDE in prognosis, tumor immunity, and therapeutic sensitivity in low grade glioma patients

BackgroundColorectal tumor differentially expressed (CRNDE) is specifically expressed in human brains and is the most highly expressed lncRNA in gliomas. Nevertheless, its implications in low grade glioma (LGG) are still indistinct. This study presented systematic analyses of CRNDE in LGG biology.MethodsWe retrospectively retrieved TCGA, CGGC and GSE16011 LGG cohorts. Survival analysis was conducted for evaluating the prognostic significance of CRNDE in LGG. A CRNDE-based nomogram was established, and its predictive performance was verified. Signaling pathways underlying CRNDE were analyzed through ssGSEA and GSEA approaches. The abundance of immune cells and activity of cancer-immunity cycle were estimated with ssGSEA approach. Immune checkpoints, HLAs, chemokines, and immunotherapeutic response indicators (TIDE, and TMB) was quantified. U251 and SW1088 cells were transfected with specific shRNAs of CRNDE, and flow cytometry (apoptosis) and western blot (β-catenin and Wnt5a) assays were conducted.ResultsUp-regulated CRNDE was found in LGG, and was linked to unfavorable clinical outcomes. The CRNDE-based nomogram enabled to accurately predict patients’ prognosis. High CRNDE expression was linked to more genomic variations, activity of tumorigenic pathways, tumor immunity (increase in infiltration of immune cells, expression of immune checkpoints, HLAs and chemokines, and cancer-immunity cycle), and therapeutic sensitivity. CRNDE knockdown mitigated malignant phenotypes of LGG cells.ConclusionsOur study determined CRNDE as a novel predictor for patient prognosis, tumor immunity and therapeutic response in LGG. Assessment of CRNDE expression is a promising approach for predicting the therapeutic benefits of LGG patients.

Relationship between gut microbiome Fusobacterium nucleatum and LINE-1 methylation level in esophageal cancer.

We previously demonstrated the relationship of human microbiome Fusobacterium nucleatum with unfavorable clinical outcomes and inferior chemotherapeutic responses in esophageal cancer. Global DNA methylation is associated with the occurrence and development of various cancers. In our previous study, LINE-1 hypomethylation (i.e., global DNA hypomethylation) was associated with a poor prognosis in esophageal cancer. As the gut microbiota may play crucial roles in the DNA methylation of host cells, we hypothesized that F. nucleatum might influence LINE-1 methylation levels in esophageal cancer. We qualified the F. nucleatum DNA using a quantitative PCR assay and LINE-1 methylation via a pyrosequencing assay using formalin-fixed paraffin-embedded specimens from 306 esophageal cancer patients. Intratumoral F. nucleatum DNA was detected in 65 cases (21.2%). The LINE-1 methylation scores ranged from 26.9 to 91.8 (median = 64.8) in tumors. F. nucleatum DNA was related to the LINE-1 hypomethylation of tumor lesions in esophageal cancer (P < 0.0001). The receiver operating characteristic curve analysis showed that the area under the curve was 0.71 for F. nucleatum positivity. Finally, we found that the impact of F. nucleatum on clinical outcomes was not modified by LINE-1 hypomethylation (P for interaction = 0.34). F. nucleatum alters genome-wide methylation levels in cancer cells, which may be one of the mechanisms by which F. nucleatum affects the malignant behavior of esophageal cancer.

Body composition, lipid profile and clinical parameters are predictors of prognosis in patients with heart failure: Two-year follow-up

Heart failure (HF) is a complex syndrome that leads to changes in body composition and eventually results in unfavorable outcomes. This study aimed to evaluate body composition, lipid profiles and clinical parameters of patients with HF, and their associations with both survival and unfavorable clinical outcomes. This prospective cohort study included 94 adults and older people with HF. Body composition was assessed by bioelectrical impedance analysis (BIA). Anthropometric variables and lipid profile were also evaluated. Electronic medical records were checked to collect information on clinical outcomes (mortality and hospitalization), considering a follow-up period of 24 months. Survival was calculated using the Kaplan-Meier estimate, and the curves compared using Log-Rank. The death risk rate (Hazard Ratio, HR) was calculated using Cox's univariate models. Mean age was 55.1 (13.9) years and there was a higher frequency of males. There was a predominance of HF with reduced ejection fraction, and ischemic etiology. Patients with New York Heart Association (NYHA) functional classification I/II had a better overall survival rate at 24 months than those with NYHA III/IV (univariate HR 4.93 (1.76-13.82); p=0.001). Greater survival rates were found in patients without chronic kidney disease (CKD) (univariate HR 2.93 (1.59-5.39); p=0.01). In the multivariate analyses, both dyslipidemia (adjusted HR 3.84 (1.22-12.00); p=0.021) and increased fat mass index (FMI) were associated with overall survival rate (adjusted HR 3.59 (1, 10-11.74); p=0.034). The severity of HF symptoms and the presence of chronic kidney disease are associated with higher mortality. Increased fat mass index and dyslipidemia are predictors of favorable outcomes in this population.

"To be or not to Be": Regulatory T cells in melanoma.

In spite of progresses in the therapy of different malignancies, melanoma still remains as one of lethal types of skin tumor. Melanoma is almost easily treatable by surgery alone with higher overall survival rates when it is diagnosed at early stages. However, survival rates are decreased remarkably upon survival if the tumor is progressed to advanced metastatic stages. Immunotherapeutics have been prosperous in the development of anti-tumor responses in patients with melanoma through promotion of the tumor-specific effector T cells in vivo; nonetheless, suitable clinical outcomes have not been satisfactory. One of the underlying causes of the unfavorable clinical outcomes might stem from adverse effects of regulatory T (Treg) cell, which is a prominent mechanism of tumor cells to escape from tumor-specific immune responses. Evidence shows that a poor prognosis and low survival rate in patients with melanoma can be attributed to a higher Treg cell number and function in these subjects. As a result, to promote melanoma-specific anti-tumor responses, depletion of Treg cells appears to be a promising approach; even though the clinical efficacy of different approaches to attain appropriate Treg cell depletion has been inconsistent. Here in this review, the main purpose is to assess the role of Treg cells in the initiation and perpetuation of melanoma and to discuss effective strategies for Treg cell modulation with the aim of melanoma therapy.

Serial measurements of lymphocytes, D-dimer, LDH, and CRP do not improve the ability to predict COVID-19 adverse outcomes

We evaluated the discriminatory ability of variations in lymphocyte, D-dimer, C-reactive protein (CRP), and lactate dehydrogenase (LDH) serum levels at 48 to 72 hours of hospitalization compared with baseline measurements to predict unfavorable clinical outcomes in patients with COVID-19. We analyzed diagnostic test results based on a retrospective cohort to determine the ability of variations (gradients or ratios) in patients' lymphocyte, D-dimer, CRP, and LDH serum levels taken 48 to 72 hours after hospital admission to predict adverse outcomes such as death, mechanical ventilation, or intensive care unit (ICU) admission developing. Among 810 patients (56.1% men, age 61.6 ± 16.2 years), 37.5% had at least one adverse outcome; 28.2% required ICU admission, 26.5% required mechanical ventilation, and 19.4% died during hospitalization. In comparing baseline measurements with measurements at 48 to 72 hours, D-dimer, lymphocyte delta, LDH, and CRP had similar discriminatory ability (area under the receiver operating characteristic curve [AUC] 0.57 vs. 0.56, 0.53 vs. 0.57, 0.64 vs. 0.66, and 0.62 vs. 0.65, respectively). Measuring serum risk markers upon hospital admission can be used to evaluate risk of adverse outcomes in hospitalized patients with COVID-19. Repeating these measurements at 48 to 72 hours does not improve discriminatory ability.

Optimal management of patients after acute coronary syndrome.

Patients with clinically established atherosclerotic cardiovascular disease are at a very high risk of recurrent cardiovascular events. An adequate management of risk factors and the implementation of healthy behaviours significantly decrease the risk of unfavourable clinical outcomes and future cardiovascular events, including death. Patients discharged after an acute coronary syndrome should be managed according to their individual risk level in order to ensure the appropriate treatment. Nevertheless, care pathways should also take into consideration the available resources and the logistical/structural aspects. In this setting, cardiac rehabilitation is prosed as a multidisciplinary approach to improving daily function and reducing cardiovascular risk factors. The organization of a network with the involvement of different medical and non-medical figures is essential to ensure successful outcomes and expected cost-effectiveness.

Pharmacoeconomics of novel pharmacotherapies in triple-negative breast cancer

ABSTRACT Introduction Triple-negative breast cancer (TNBC) represents the most aggressive breast cancer subtype carrying unfavorable clinical outcomes. Although traditionally chemotherapy has represented the only systemic treatment option available, novel drugs have changed the treatment landscape, granting approval by regulatory agencies worldwide, while determining new economic struggles on health systems for their high prices. Areas covered In this review, we provided a comprehensive analysis of pharmacoeconomic studies of drugs recently approved in the early and advanced settings of TNBC. Expert opinion Novel systemic treatment options redefined the therapeutic algorithms for TNBC by establishing new paradigms, based on substantial clinical benefits. Pembrolizumab and olaparib in the curative setting portend high value, as shown with the use of value frameworks, resulting in cost-effective interventions. In the metastatic setting, new drugs have demonstrated mixed improvements in patient-centric end-points, resulting often in interventions unlikely to have good value for money. We believe that cost-effectiveness alone is not a metric to inform the opportunity to invest on new interventions, while the intrinsic value of medicines should be the driver of the decisions. We endorse a patient-centric priority setting that can ensure health system sustainability, to timely deliver innovative cancer care to all in need and positively impact on population health.

A pilot study: Metabolic profiling of plasma and saliva samples from newly diagnosed glioblastoma patients.

Despite aggressive treatment, more than 90% of glioblastoma (GBM) patients experience recurrences. GBM response to therapy is currently assessed by imaging techniques and tissue biopsy. However, difficulties with these methods may cause misinterpretation of treatment outcomes. Currently, no validated therapy response biomarkers are available for monitoring GBM progression. Metabolomics holds potential as a complementary tool to improve the interpretation of therapy responses to help in clinical interventions for GBM patients. Saliva and blood from GBM patients were collected pre and postoperatively. Patients were stratified conforming their progression-free survival (PFS) into favourable or unfavourable clinical outcomes (>9months or PFS ≤ 9months, respectively). Analysis of saliva (whole-mouth and oral rinse) and plasma samples was conducted utilising LC-QqQ-MS and LC-QTOF-MS to determine the metabolomic and lipidomic profiles. The data were investigated using univariate and multivariate statistical analyses and graphical LASSO-based graphic network analyses. Altogether, 151 metabolites and 197 lipids were detected within all saliva and plasma samples. Among the patients with unfavourable outcomes, metabolites such as cyclic-AMP, 3-hydroxy-kynurenine, dihydroorotate, UDP and cis-aconitate were elevated, compared to patients with favourable outcomes during pre-and post-surgery. These metabolites showed to impact the pentose phosphate and Warburg effect pathways. The lipid profile of patients who experienced unfavourable outcomes revealed a higher heterogeneity in the abundance of lipids and fewer associations between markers in contrast to the favourable outcome group. Our findings indicate that changes in salivary and plasma metabolites in GBM patients can potentially be employed as less invasive prognostic biomarkers/biomarker panel but validation with larger cohorts is required.

Study the Relationship Physiology and Molecular Biology in Patients of Iraq Drug Use Parameters Acute Leukaemia (AL)

Study of paper is physiology use to analysis total in LDH is an intracellular enzyme whose activity is associated with cellular metabolism, WBCs count in blood to patients which makes it a marker of cell proliferation and cell injury and study with molecular biology of marker parameters use drug the presence of survivin in neoplastic cells contributes to tumor development and its high transcription has been reported as a predictor of poor prognosis in several hematological malignancies. However, despite being frequently present in ALs, the impact of survivin transcription in this type of hematologic malignancy remains undefined. Some studies report that survivin predicts an unfavorable clinical outcome in ALs, while others do not. In the present study, despite the small number of patients evaluated, the transcription of survivin at diagnosis did not seem to be related to a worse response to treatment or a worse prognosis. This is because, of the six patients who expressed survivin, three are in complete remission and, of the three who died, two were already considered to have an unfavorable prognosis regardless of survivin transcription, one due to advanced age and the presence of leukocytosis in the liver. Leukometry at diagnosis was another prognostic factor evaluated in this study. Patients diagnosed with AL may have elevated, normal, or decreased WBC counts at the time of diagnosis and, in general, counts greater than 30,000 WBCs/mm3 are considered to have a poor prognosis.

The clinicopathologic significance of NPM1 mutation and ability to detect mutated NPM1 by immunohistochemistry in non-AML myeloid neoplasms.

NPM1 mutated non-AML myeloid neoplasms (MN; <20% blasts) are characterized by an aggressive clinical course in a few studies. In this retrospective study, we evaluate the clinicopathologic and immunohistochemical features of non-AML MN patients with NPM1 mutations. We assessed NPM1 mutation by targeted next generation sequencing (NGS). Cytoplasmic NPM1 expression was assessed by immunohistochemistry (IHC) on formalin-fixed, formic acid-decalcified bone marrow biopsy specimens. We evaluated 34 non-AML MN patients with NPM1 mutations comprising MDS (22), MPN (3) and MDS/MPN (9). They commonly presented with anemia (88%), thrombocytopenia (58%) and leukopenia (50%). Bone marrow dysplasia was common (79%). The karyotype was often normal (64%). NGS for MN-associated mutations performed in a subset of the patients showed a median of 3 mutations. NPM1 mutations were more often missense (c.859C > T p. L287F; 65%) than frameshift insertion/duplication (35%) with median variant allele frequency (VAF; 9.7%, range 5.1%-49.8%). Mutated NPM1 by IHC showed cytoplasmic positivity in 48% and positivity was associated with higher VAF. The median overall survival (OS) in this cohort was 70 months. Nine patients (26%) progressed to AML. OS in patients who progressed to AML was significantly shorter than the one of patients without progression to AML (OS 20 vs. 128 months, respectively, log rank p = 0.05). NPM1 mutated non-AML MN patients commonly had cytopenias, dysplasia, normal karyotype, mutations in multiple genes, and an unfavorable clinical outcome, including progression to AML. Our data demonstrated that IHC for NPM1 can be a useful supplementary tool to predict NPM1 mutation in some non-AML MN; however, genetic testing cannot be replaced by IHC assessment.

Abstract 3264: Sotorasib, a selective G12C K-RAS inhibitor, modulates the tumor microenvironment to promote TNFα and IFNγ signaling and augment immunotherapy response

Abstract KRAS-G12C mutation represents a highly prevalent mutation in lung and colon carcinoma with unfavorable clinical outcome. Recently, new compounds were developed that selectively bind to K-RAS with G12C mutation. While recent clinical results demonstrate activity of these compounds, their underlying mechanisms of action and impact on the antitumor immune response are not well understood. Here, we tested G12C-Kras inhibitor (Sotorasib/AMG-510) using multiple approaches. We found that Sotorasib significantly promoted the tumor regression and the expansion of tumor infiltrating lymphocytes (TILs) in both CT26-G12C heterotopic colon cancer, and KP-G12C autochthonous GEMM lung cancer mouse models, with more profound impact when combined with anti-PD1 ICB immunotherapy. In addition, it generated a “hot” tumor microenvironment with strikingly augmented T cell effector phenotype, endorsed by enrichment of MHC-II high macrophage and reduction of suppressive myeloid cells. Interestingly, Single cell RNA sequencing of CT26-G12C tumors shows an enrichment of tumor clusters with TNFα and IFNγ as top upregulated pathways in Sotorasib-treated tumors, generating a more immune active TME. These immune active tumor clusters were conserved or further expanded upon cotreatment with anti-PD1 ICI. Conversely, immune-deficient, or Sotorasib-resistant tumors exhibited a remarkable contraction of the immune active clusters, substantiating that the immune-modulatory impact of Sotorasib relies on tumor cells adaptation and their cross talk with the tumor infiltrated lymphocytes. In vitro, Single cell RNA sequencing of CT26-G12C cells also revealed that Sotorasib and TNFα/IFNγ cytokines are affecting different tumor clusters. Moreover, further in vitro testing demonstrated that Sotorasib dramatically enhances KRAS-G12C mutant cancer cell killing by PSCA-specific CART cells. These findings provide insight into the molecular basis of G12C-Kras inhibitor-mediated immune modulation in the TME and provide insight for combinatorial regimens with checkpoint inhibitor and adoptive T cell immunotherapy to achieve optimum anti-tumor activity. Citation Format: Mohammed L. Ibrahim, Dana Foresman, Hong Zheng, Eric B. Haura, Daniel Abate-Daga, Xiaoqing Yu, Amer A. Beg. Sotorasib, a selective G12C K-RAS inhibitor, modulates the tumor microenvironment to promote TNFα and IFNγ signaling and augment immunotherapy response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3264.

Abstract 1555: CDK2 inhibition &amp; disruption of centrosome stoichiometry: a clinically tractable death program for aneuploid cancer cells

Abstract Genomic instability is a cancer hallmark. Aberrant centrosome numbers often occur in cancer and is associated with genomic instability. Mitosis with supernumerary centrosomes leads to abnormal chromosome segregation and aneuploidy in progeny cells. While many cells with abnormal chromosomes are eliminated through apoptosis, some gain a growth advantage and acquire resistance to antineoplastic agents. This aneuploidy is associated with aggressive tumor biology and unfavorable clinical outcomes. Cyclin-dependent kinase 2 (CDK2) plays a major role regulation of cell cycle progression, centrosome duplication and clustering. We previously reported that CDK2 inhibition prevented supernumerary centrosomes clustering, causing multipolar mitosis and anaphase catastrophe in lung and other aneuploid cancers. To elucidate engaged mechanisms focused ion beam scanning electron microscope (FIB-SEM) and immunofluorescent staining were each used to interrogate the ultrastructure of multipolar mitosis after CDK2 inhibition by CYC065-treatment (a CDK2/9 inhibitor). CYC065-treatment induced distinct ultrastructural changes in aneuploid lung cancer cells including formation of chromosome rings and multipolarity. As compared to vehicle controls, CYC065-treatment markedly extended the duration of chromosome rings in lung cancer cells and conferred aberrant mitosis in live cell imagining. Intriguingly, this treatment disrupted centrosome and centriole stoichiometry in cells with chromosome rings or multipolarity. These findings were confirmed using immunofluorescence assays of lung and other cancer cells having supernumerary centrosomes. To elucidate if CDK2 or CDK9 activity directly altered centriole stoichiometry, chromosome rings or multipolarity, immunofluorescent assays were performed in lung cancer cells that independently knocked down CDK2 or CDK9 using siRNAs relative to controls. Centriole stoichiometry abnormalities in cancer cells having chromosome rings or multipolarity were statistically significantly higher in CDK2 knock-down but not in CDK9 knock-down transfectants or controls. Engineered gain of CDK2 but not CDK9 expression rescued CYC065 mediated chromosome rings and multipolarity in aneuploid lung cancer cells. Taken together, this study uncovered that CDK2 inhibition disrupted centrosome stoichiometry, causing apoptotic death of aneuploid cancer cells. This is a clinically-tractable pharmacologic mechanism that conferring cancer cell death. Current work determines if similar mechanisms are engaged in vivo in syngeneic mouse lung cancer models undergoing CDK2 antagonism. Citation Format: Zibo Chen, Xi Liu, Masanori Kawakami, Xiuxia Liu, Allison Baker, Aayush Bhatawadekar, Liliya Tyutyunyk-Massey, Ethan Dmitrovsky, Kedar Narayan. CDK2 inhibition &amp; disruption of centrosome stoichiometry: a clinically tractable death program for aneuploid cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1555.