Association of tumor budding with clinicopathological features and prognostic value in stage III-IV colorectal cancer

BACKGROUND Tumor budding(TB)has emerged as a promising independent prognostic biomarker in colorectal cancer(CRC).The prognostic role of TB has been extensively studied and currently affects clinical decision making in patients with stage I and II CRC.However,existing prognostic studies on TB in stage III CRC have been confined to small retrospective cohort studies.Consequently,this study investigated the correlation among TB categories,clinicopathological features,and prognosis in stage III-IV CRC to further enhance the precision and individualization of treatment through refined prognostic risk stratification.AIM To analyze the relationship between TB categories and clinicopathological characteristics and assess their prognostic value in stage III-IV CRC to further refine the prognostic risk stratification of stage III-IV CRC.METHODS The clinical data of 547 CRC patients were collected for this retrospective study.Infiltration at the front edge of the tumor buds was counted according to the 2016 International Tumor Budding Consensus Conference guidelines.RESULTS Multivariate Cox proportional hazards regression analysis demonstrated that chemotherapy(P=0.004),clinical stage IV(P<0.001),≥4 regional lymph node metastases(P=0.004),left-sided colonic cancer(P=0.040),and Bd 2-3(P=0.002)were independent prognostic factors in patients with stage III-IV CRC.Moreover,the density of tumor infiltrating lymphocytes was higher in Bd 1 than in Bd 2-3,both in the tumor stroma and its invasive margin.CONCLUSION TB has an independent predictive prognostic value in patients with stage III-IV CRC.It is recommended to complete the TB report of stage III-IV CRC cases in the standardized pathological report to further refine risk stratification.

Mathematical Modeling of Cell Polarity Establishment of Budding Yeast

The budding yeast Saccharomyces cerevisiae is a powerful model system for studying the cell polarity establishment.The cell polarization process is regulated by signaling molecules,which are initially distributed in the cytoplasm and then recruited to a proper location on the cell membrane in response to spatial cues or spontaneously.Polarization of these signaling molecules involves complex regulation,so the mathematical models become a useful tool to investigate the mechanism behind the process.In this review,we discuss how mathematical modeling has shed light on different regulations in the cell polarization.We also propose future applications for the mathematical modeling of cell polarization and morphogenesis.

Prognostic significance of tumor budding,desmoplastic reaction,and lymphocytic infiltration in patients with gastric adenocarcinoma

BACKGROUND Recent studies have shown that the tumor microenvironment significantly influences the behavior of solid tumors.In this context,Accumulated data suggests that pathological evaluation of tumor budding(TB),desmoplastic reaction(DR),and tumor-infiltrating lymphocytes(TILs)may be crucial in determining tumor behavior in the gastrointestinal tract.Regarding gastric adenocarcinoma(GAC),although some results suggest that TB and TILs may be effective in determining the course of the disease,the data do not agree.Moreover,very few studies have investigated the relationship between DR and survival.At present,the associations between tumor TB,DR and TILs in GAC patients have not been determined.AIM To establish the relationships between TB,DR,and TILs in patients with GAC and to assess their influence on prognosis.METHODS Our study group comprised 130 patients diagnosed with GAC.The definition of TB was established based on the International TB Consensus Conference.The DR was categorized into three groups according to the level of tumor stroma maturation.The assessment of TILs was conducted using a semiquantitative approach,employing a cutoff value of 5%.The statistical analysis of the whole group and 100 patients with an intestinal subtype of GAC was performed using SPSS version 27.RESULTS A significant correlation between peritumoral budding(PTB)and intratumoral budding(ITB)was noted(r=0.943).Tumors with high PTBs and ITBs had a greater incidence of immature DRs and low TILs(P<0.01).PTB and ITB were associated with histological subtype,lymph node metastasis(LNM),and stage(P<0.01).ITB,PTB,LNM,DR,and stage were significant risk factors associated with poor prognosis.The multivariate Cox regression analysis identified ITB,PTB,and LNM as independent prognostic variables(P<0.05).In intestinal-type adenocarcinomas,a positive correlation between PTB and ITB was noted(r=0.972).While univariate analysis revealed that LNM,stage,PTB,ITB,and DR were strong parameters for predicting survival(P<0.05),only PTB and ITB were found to be independent prognostic factors(P<0.001).CONCLUSION TB may be a potential prognostic marker in GAC.However,further studies are needed to delineate its role in pathology reporting protocols and the predictive effects of DR and TILs.

Artificial intelligence-based comprehensive analysis of immune-stemness-tumor budding profile to predict survival of patients with pancreatic adenocarcinoma

Objective:Pancreatic ductal adenocarcinoma(PDAC)is an aggressive malignancy.CD8^(+)T cells,cancer stem cells(CSCs),and tumor budding(TB)have been significantly correlated with the outcome of patients with PDAC,but the correlations have been independently reported.In addition,no integrated immune-CSC-TB profile for predicting survival in patients with PDAC has been established.Methods:Multiplexed immunofluorescence and artificial intelligence(AI)-based comprehensive analyses were used for quantification and spatial distribution analysis of CD8^(+)T cells,CD133^(+)CSCs,and TB.In vivo humanized patient-derived xenograft(PDX)models were established.Nomogram analysis,calibration curve,time-dependent receiver operating characteristic curve,and decision curve analyses were performed using R software.Results:The established‘anti-/pro-tumor’models showed that the CD8^(+)T cell/TB,CD8^(+)T cell/CD133^(+)CSC,TB-adjacent CD8^(+)T cell,and CD133^(+)CSC-adjacent CD8^(+)T cell indices were positively associated with survival of patients with PDAC.These findings were validated using PDX-transplanted humanized mouse models.An integrated nomogram-based immune-CSC-TB profile that included the CD8^(+)T cell/TB and CD8^(+)T cell/CD133^(+)CSC indices was established and shown to be superior to the tumor-nodemetastasis stage model in predicting survival of patients with PDAC.Conclusions:‘Anti-/pro-tumor’models and the spatial relationship among CD8^(+)T cells,CSCs,and TB within the tumor microenvironment were investigated.Novel strategies to predict the prognosis of patients with PDAC were established using AI-based comprehensive analysis and machine learning workflow.The nomogram-based immune-CSC-TB profile can provide accurate prognosis prediction for patients with PDAC.

Tumor budding in gastric cancer

The tumor,nodes,metastasis(TNM)staging system has long been the gold standard for the classification and prognosis of solid tumors.However,the TNM staging system is not without limitations.Prognostic heterogeneity exists within patients at the same stage.Therefore,the pursuit of other biomarkers with the potential to classify patients with cancer has never stopped.One of them,tumor budding(TB),has gained much success in colorectal cancer.In recent years,TB in gastric cancer has attracted much attention from researchers,beginning to reveal the molecular and biological aspects of this phenomenon in gastric cancer,and has emerged as a promising prognostic biomarker in gastric cancer,predicting disease progression and unfavorable survival.Therefore,it is time and essential to provide a holistic overview of TB in gastric cancer,which has not been achieved and is the aim of this review.

Tumor budding as a potential prognostic marker in determining the behavior of primary liver cancers

Hepatocellular(HCC)and intrahepatic cholangiocarcinoma(ICC),the most common primary tumors of the liver,are among the most important causes of cancer deaths worldwide.Because patients with primary liver tumors are frequently diagnosed at an advanced stage and have high mortality,many efforts have been made to identify new markers to determine their behavior and treatment,similar to those in other solid organ tumors.Recently,morphological assessment of tumor budding(TB)has been revealed as a promising prognostic finding to predict tumor behavior and survival across several different tumor types.Currently,the TB score in colorectal cancer has been revealed as an important parameter in pathology report protocols to determine the course of the disease.Regarding the liver,despite enormous data showing that many mechanisms involved in TB are associated with tumor behavior in both HCC and ICC,studies focusing on the role of TB in predicting the behavior and prognosis of these tumors have started to be investigated very recently.The purpose of this review is to present data about TB in primary tumors of the liver,pointing out the potential role of this parameter in determining the course of the disease,and emphasize the need to increase the number of further studies focusing on the evaluation of this parameter with an overview of the mechanisms involved in TB.

Parasympathetic neurogenesis is strongly associated with tumor budding and correlates with an adverse prognosis in pancreatic ductal adenocarcinoma

Objective： To investigate the frequency of parasympathetic neurogenesis and determine its association with tumor budding and prognosis in pancreatic ductal adenocarcinoma （PDAC）. Methods： Parasympathetic neurogenesis was defined as the distribution of abnormal parasympathetic nerves in the stroma tissue. Staining of vesicular acetylcholine transporter （VAChT）, as a marker for parasympathetic neurogenesis, was performed on a representative specimen of the tumor for 59 PDAC patients with available clinical, pathologic, and follow-up information. Three specimens containing normal pancreatic tissues were stained in parallel. The number of parasympathetic nerve fibers was counted in five high-power microscopic fields （5×0.785 mm2）. Cut-offvalues were calculated by receiver operating characteristic curve analysis. Results： VAChT-positive parasympathetic nerve fibers were not seen in the stroma of 3 cases of normal pancreatic tissues. In 59 PDAC cases, the range of parasympathetic neurogenesis was 4-38 fibers/（5×0.785） mm2, with a median of 18 fibers/（5×0.785） mm2. Patients with parasympathetic neurogenesis 〉 15 fibers/（5×0.785） mm2 were defined as the high-density group （39 patients, 66.1%）, and those with parasympathetic neurogenesis 〈15 fibers/（5×0.785） mm2 as the low-density group （20 patients, 33.9%）. The high-density group had a higher occurrence of tumor budding （P=0.001） and a higher rate of early recurrence （P=0.035）. Parasympathetic neurogenesis appeared to be an independent adverse prognostic factor [hazard ratio （HR）=2.45, 95% confidence interval （95% CI）： 1.25-4.81, P=0.009], in addition to American Joint Committee on Cancer （AJCC） stage （P=0.010） and tumor budding （P=0.009）. Conclusions： Parasympathetic neurogenesis is strongly associated with tumor budding and correlates with an adverse prognosis in PDAC.

Tumor budding predicts response to anti-EGFR therapies in metastatic colorectal cancer patients

AIM:To investigate whether the evaluation of tumor budding can complement K-RAS analysis to improve the individualized prediction of response to anti-epidermal growth factor receptor based therapies in metastatic colorectal cancer (mCRC) patients. METHODS:Forty-three patients with mCRC treated with cetuximab or panitumumab were entered into this study. According to the Response Evaluation Criteria in Solid Tumors criteria, 30 patients had stable or progressive disease (non-responsive), while 13 patients had a partial response. Tumor buds were evaluated from whole tissue sections stained for pan-cytokeratin, evaluated in the densest region using a 40 × objective and "high-grade" tumor budding was defi ned as 15 buds/high-power f ield.RESULTS: Tumor buds and K-RAS mutation both correctly classif ied 68% of patients. All patients with K-RAS mutation (n=7) or high-grade tumor budding (n=11) were non-responsive, of which 4 patients had both features. All 13 partial responders were K-RAS wild-type with low-grade tumor budding. Combined, the predictive value of K-RAS and tumor budding was 80%. Additionally, high-grade tumor budding was significantly related to worse progression-free survival [HR (95% CI): 2.8 (1.3-6.0, P=0.008)].CONCLUSION: If confirmed in larger cohorts, the addition of tumor budding to K-RAS analysis may represent an effective approach for individualized patient management in the metastatic setting.

Prognostic and pathological impact of tumor budding in gastric cancer: A systematic review and meta-analysis

BACKGROUND Tumor budding,is a promising prognostic hallmark in many cancers,and can help us better assess the degree of malignancy in gastric cancer(GC)and in colorectal cancer.In the past few years,several articles on the relationship between tumor budding and GC have been published,but different results have been observed.As the relationship between tumor budding and GC remains controversial,we integrated the data from 7 eligible studies to conduct a systematic review and meta-analysis.AIM To systematically evaluate the prognostic and pathological impact of tumor budding in GC.METHODS Literature searches were conducted in the PubMed,Cochrane Library,EMBASE and Web of Science databases,and 7 cohort studies involving 2178 patients met our criteria and included in the analysis.The patients were divided into those with high-grade tumor budding and those with low-grade tumor budding,and the cut-off values for tumor budding varied across the included studies.The hazard ratios(HRs)with 95%confidence intervals(CIs)were calculated to estimate the impact of tumor budding on overall survival(OS)in GC patients.The odds ratios(ORs)with 95%CIs were used to determine the correlation between tumor budding and pathological parameters(tumor stage,tumor differentiation,lymphovascular invasion,lymph node metastasis)of GC.RESULTS Seven studies involving 2178 patients were included in the meta-analysis.The combined ORs suggested that high-grade tumor budding was significantly associated with tumor stage(OR=6.63,95%CI:4.01-10.98,P<0.01),tumor differentiation(OR=3.74,95%CI:2.68-5.22,P<0.01),lymphovascular invasion(OR=7.85,95%CI:5.04-12.21,P<0.01),and lymph node metastasis(OR=5.75,95%CI:3.20-10.32,P<0.01).Moreover,high-grade tumor budding predicted a poor 5-year OS(HR=1.79,95%CI:1.53-2.05,P<0.01)in patients with GC and an adverse 5-year OS(HR=1.93,95%CI:1.45-2.42,P<0.01)in patients with intestinal-type GC.CONCLUSION High-grade tumor budding suggested a poor prognosis in patients with GC or intestinal-type GC.

Prognostic role of tumor budding in breast cancer

Tumor budding, defined as a small number of cancer cells observed in pathology sections detached from the main tumor mass, is a common phenomenon in cancer. It issuggested that cells in buds are in the process of actively moving away from the primary tumor in the first step of metastasis. Tumor budding has been observed in a variety of carcinomas and is best studied in colorectal cancers where it portends poor prognosis. More recently, tumor budding was found to be of prognostic significance in other cancers including breast cancer. Tumor budding in breast cancer is associated with other adverse pathologic factors, such as larger tumor size and lymphovascular invasion, but may have additional independent prognostic value. In the future, standardization of the quantification criteria for tumor budding may further aid in its adoption as a prognostic marker.

Mismatch repair protein expression and intratumoral budding in rectal cancer are associated with an increased pathological complete response to preoperative chemoradiotherapy: A case-control study

AIM To determine whether the association of rectal adenocarcinoma with a defective-mismatch repair system(dMMR) was associated with a pathological complete response(pCR) to preoperative chemoradiotherapy.METHODS A case-control study was designed with the aim of determining if patients with rectal adenocarcinoma with dM MR had an associated high pCR rate in response to neoadjuvant chemoradiotherapy(nCRT).RESULTS Seventy-two cases with pCR were compared against 144 controls without pCR. Across 216 cases, the mean age was 56.8 years, 140(64.8%) were men, and 63(29.2%) demonstrated the dMMR system. The pCR was associated with G1 tumors, dMMR, the absence of vascular invasion, and low tumor budding in the pretreatment biopsy. In a multivariant analysis, the factors associated with pCR were dMMR(OR: 2.61; 95%CI: 1.355-5.040, P = 0.004) and a low degree of tumor budding(OR: 2.52; 95%CI: 1.366-4.894, P = 0.025).CONCLUSION We found an independent association between dMMR and a low rate of tumor budding, with a higher rate of pCR, in the basal biopsies of patients with rectal carcinoma subjected to nCRT.

Circulating cytokeratin-positive cells and tumor budding in colorectal cancer

AIM To investigate whether circulating cytokeratin-positive(CK^+) cells in the mesenteric blood of resected colorectal specimens are prognostic and correlate with tumor budding.METHODS Fifty-six colorectal specimens were collected between 9/2007 and 7/2008.Blood from the mesenteric vein was drawn immediately after receiving the fresh and unfixed specimens in the pathology department.After separation of the mononuclear cells by Ficoll-Hypaquedensity-gradient centrifugation,cytological smears were immunocytochemically stained for CK18.Tumor budding was evaluated on slides stained for pan-cytokeratin.The identification of ≥ 30 buds/1.3 mm2 was defined as high grade budding.RESULTS CK^+ cells and clusters were identified in 29(48%) and 14(25%) of the samples,respectively.Two cells were identified in one of three non-malignant cases.Clusters were found exclusively in malignant cases.The occurrence of CK^+ cells or clusters was not associated with any of the evaluated clinicopathological factors,including surgical technique and tumor budding.Moreover,the occurrence of CK^+ cells or clusters had no influence on the cancerspecific survival [75 mo(CI:61;88) vs 83 mo(CI:72;95) and 80 mo(CI:63;98) vs 79 mo(CI:69;89),respectively].CONCLUSION CK^+ cells and showed neither prognostic significance nor an association with tumor budding.It is very likely that CK18-staining is not specific enough to identify the relevant cells.

Role of Self-Loop in Cell-Cycle Network of Budding Yeast

Study of network dynamics is very active area in biological and social sciences. However, the relationship between the network structure and the attractors of the dynamics has not been fully understood yet. In this study, we numerically investigated the role of degenerate self-loops on the attractors and its basin size using the budding yeast cell-cycle network model. In the network, all self-loops negatively suppress the node (self-inhibition loops) and the attractors are only fixed points, i.e. point attractors. It is found that there is a simple division rule of the state space by removing the self-loops when the attractors consist only of point attractors. The point attractor with largest basin size is robust against the change of the self-inhibition loop. Furthermore, some limit cycles of period 2 appear as new attractor when a self-activation loop is added to the original network. It is also shown that even in that case, the point attractor with largest basin size is robust.

Mec1-Dependent Phosphorylation of the Scc3 Subunit of Cohesin during Mitosis in Budding Yeast

Cohesin is an evolutionary conserved complex that controls chromosome segregation during mitosis. Here we show that, in response to DNA damage, Saccharomyces cerevisiae Scc3, one of the major regulatory subunits of the Smc1-Smc3-Scc1 cohesin ring, is phosphorylated on S/T-Q residues. This event depended on the Mec1 checkpoint kinase as well as on cell cycle arrest triggered by the DNA damage checkpoint network. This phosphorylation event also took place during mitosis of an unperturbed cell cycle. The present finding that S. cerevisiae Scc3 is phosphorylated during mitosis represents a potentially important new regulatory step in cohesin’s mitotic functions.

Research Progress on Molecular Mechanisms of Tumor Budding in Colorectal Cancer

Tumor buds are usually defined as isolated single cancer cells or clusters of up to four cancer cells located at the front of invasive tumors which play an important role in the clinical pathological study of colorectal cancer.The prognostic value of tumor budding in colorectal cancer has been supported by a large amount of evidence.However,its molecular mechanism remains unclear,and it is also a research hotspot now.This paper reviews the latest research progress on the molecular mechanisms of tumor budding in colorectal cancer.

Risk factors for lymph node metastasis in superficial esophageal squamous cell carcinoma

In this editorial,we comment on the article by Wang et al published in the recent issue of the World Journal of Gastroenterology in 2023.We focused on identifying risk factors for lymph node metastasis(LNM)in superficial esophageal squamous cell carcinoma(SESCC)patients and how to construct a simple and reliable clinical prediction model to assess the risk of LNM in SESCC patients,thereby helping to guide the selection of an appropriate treatment plan.The current standard treatment for SESCC is radical esophagectomy with lymph node dissection.However,esophagectomy is associated with considerable morbidity and mortality.Endoscopic resection(ER)offers a safer and less invasive alternative to surgical resection and can enable the patient's quality of life to be maintained while providing a satisfactory outcome.However,since ER is a localized treatment that does not allow for lymph node dissection,the risk of LNM in SESCC limits the effectiveness of ER.Understanding LNM status can aid in determining whether patients with SESCC can be cured by ER without the need for additional esophagectomy.Previous studies have shown that tumor size,macroscopic type of tumor,degree of differentiation,depth of tumor invasion,and lymphovascular invasion are factors associated with LNM in patients with SESCC.In addition,tumor budding is commonly associated with LNM,recurrence,and distant metastasis,but this topic has been less covered in previous studies.By comprehensively evaluating the above risk factors for LNM,useful evidence can be obtained for doctors to select appropriate treatments for SESCC patients.

Hrs inhibits citron kinase-mediated HIV-1 budding via its FYVE domain

Hepatocyte growth factor-regulated tyrosine kinase substrate(Hrs)is a key component of the endosomal sorting complexes required for transport and has been demonstrated to play a regulatory role in endocytosis/exocytosis and the accumulation of internal vesicles in multivesicular bodies.Citron kinase is a Ser/The kinase that we previously reported to enhance human immunodeficiency virus type 1(HIV-1)virion production.However,the relationship between Hrs and citron kinase in HIV-1 production remains elusive.Here,we report that Hrs interacts with citron kinase via its FYVE domain.Overexpression of Hrs or the FYVE domain resulted in a significant decrease in HIV-1 virion production.Depletion of Hrs by RNA interference in HEK293T cells increased HIV-1 virion production and enhanced the activity of citron kinase.These data suggest that Hrs inhibits HIV-1 production by inhibiting citron kinase-mediated exocytosis.

Peculiarity of transcriptional and H3K27me3 dynamics during peach bud dormancy

Bud dormancy facilitates the survival of meristems under harsh environmental conditions.To elucidate how molecular responses to chilling accumulation controlling dormancy in peach buds,chromatin immunoprecipitation sequencing to identify the H3K27me3 modifications and RNA sequencing of two peach cultivars with pronounced differences in chilling requirement were carried out,the results showed that genes associated with abscisic acid and gibberellic acid signal pathways play key roles in dormancy regulation.The results demonstrated that peach flower bud differentiation occurred continuously in both cultivars during chilling accumulation,which was correlated with the transcript abundance of key genes involved in phytohormone metabolism and flower bud development under adverse conditions.The more increased strength in high chillingrequirement cultivar along with the chilling accumulation at the genome-wide level.The function of the dormancy-associated MADS-box gene PpDAM6 was identified,which is involved in leaf bud break in peach and flower development in transgenic Nicotiana tabacum(NC89).In addition,PpDAM6 was positively regulated by PpCBF,and the genes of putative dormancy-related and associated with metabolic pathways were proposed.Taken together,these results constituted a theoretical basis for elucidating the regulation of peach bud dormancy transition.

A New Micropropagation Technology of Tilia amurensis:In VitroMicropropagation of Mature Zygotic Embryos and the Establishment of a PlantRegeneration System

Tilia amurensis is an economically valuable broadleaf tree species in Northeast China.The production of highqualityT.amurensis varieties at commercial scales has been greatly limited by the low germination rates.Thereis thus a pressing need to develop an organogenesis protocol for in vitro propagation of T.amurensis to alleviate ashortage of high-quality T.amurensis seedlings.Here,we established a rapid in vitro propagation system forT.amurensis from mature zygotic embryos and analyzed the effects of plant growth regulators and culture mediain different stages.We found that Woody plant medium(WPM)was the optimal primary culture medium formature zygotic embryos.The highest callus induction percentage(68.76%)and number of axillary buds induced(3.2)were obtained in WPM+0.89μmol/L 6-benzyladenine(6-BA)+0.46μmol/L kinetin(KT)+0.25μmol/Lindole-3-butryic acid(IBA)+1.44μmol/L gibberellin A_(3)(GA_(3)).The multiple shoot bud development achievedthe highest percentage(83.32%)in the Murashige and Skoog(MS)+2.22μmol/L 6-BA+0.25μmol/L IBA+1.44μmol/L GA_(3).The rooting percentage(96.70%)was highest in 1/2 MS medium+1.48μmol/L IBA.Thesurvival percentage of transplanting plantlets was 82.22%in soil:vermiculite:perlite(5:3:1).Our study is the firstto establish an effective organogenesis protocol for T.amurensis using mature zygotic embryos.

The two-stage interaction of Ebola virus VP40 with nucleoprotein results in a switch from viral RNA synthesis to virion assembly/ budding

Ebola virus(EBOV)is an enveloped negative-sense RNA virus and a member of the filovirus family.Nucleoprotein(NP)expression alone leads to the formation of inclusion bodies(IBs),which are critical for viral RNA synthesis.The matrix protein,VP40,not only plays a critical role in virus assembly/budding,but also can regulate transcription and replication of the viral genome.However,the molecular mechanism by which VP40 regulates viral RNA synthesis and virion assembly/budding is unknown.Here,we show that within IBs the N-terminus of NP recruits VP40 and is required for VLP-containing NP release.Furthermore,we find four point mutations(L692A,P697A,P698A and W699A)within the C-terminal hydrophobic core of NP result in a stronger VP40–NP interaction within IBs,sequestering VP40 within IBs,reducing VP40–VLP egress,abolishing the incorporation of NC-like structures into VP40–VLP,and inhibiting viral RNA synthesis,suggesting that the interaction of N-terminus of NP with VP40 induces a conformational change in the C-terminus of NP.Consequently,the C-terminal hydrophobic core of NP is exposed and binds VP40,thereby inhibiting RNA synthesis and initiating virion assembly/budding.