Evaluation Of Tumor Budding Research Articles

Evaluation of tumor budding with virtual panCK stains generated by novel multi-model CNN framework

As the global incidence of cancer continues to rise rapidly, the need for swift and precise diagnoses has become increasingly pressing. Pathologists commonly rely on H&E-panCK stain pairs for various aspects of cancer diagnosis, including the detection of occult tumor cells and the evaluation of tumor budding. Nevertheless, conventional chemical staining methods suffer from notable drawbacks, such as time-intensive processes and irreversible staining outcomes. The virtual stain technique, leveraging generative adversarial network (GAN), has emerged as a promising alternative to chemical stains. This approach aims to transform biopsy scans (often H&E) into other stain types. Despite achieving notable progress in recent years, current state-of-the-art virtual staining models confront challenges that hinder their efficacy, particularly in achieving accurate staining outcomes under specific conditions. These limitations have impeded the practical integration of virtual staining into diagnostic practices. To address the goal of producing virtual panCK stains capable of replacing chemical panCK, we propose an innovative multi-model framework. Our approach involves employing a combination of Mask-RCNN (for cell segmentation) and GAN models to extract cytokeratin distribution from chemical H&E images. Additionally, we introduce a tailored dynamic GAN model to convert H&E images into virtual panCK stains, integrating the derived cytokeratin distribution. Our framework is motivated by the fact that the unique pattern of the panCK is derived from cytokeratin distribution. As a proof of concept, we employ our virtual panCK stains to evaluate tumor budding in 45 H&E whole-slide images taken from breast cancer-invaded lymph nodes . Through thorough validation by both pathologists and the QuPath software, our virtual panCK stains demonstrate a remarkable level of accuracy. In stark contrast, the accuracy of state-of-the-art single cycleGAN virtual panCK stains is negligible. To our best knowledge, this is the first instance of a multi-model virtual panCK framework and the utilization of virtual panCK for tumor budding assessment. Our framework excels in generating dependable virtual panCK stains with significantly improved efficiency, thereby considerably reducing turnaround times in diagnosis. Furthermore, its outcomes are easily comprehensible even to pathologists who may not be well-versed in computer technology. We firmly believe that our framework has the potential to advance the field of virtual stain, thereby making significant strides towards improved cancer diagnosis.

Immunohistochemical analysis of tumor budding in stage II colon cancer: exploring zero budding as a prognostic marker

Tumor budding, a biomarker traditionally evaluated using hematoxylin and eosin (H&E) staining, has gained recognition as a prognostic biomarker for stage II colon cancer. Nevertheless, while H&E staining offers valuable insights, its limitations prompt the utilization of pan-cytokeratin immunohistochemistry (IHC). Consequently, this study seeks to evaluate the prognostic significance of tumor budding using IHC in a contemporary cohort of stage II colon cancer patients, aiming to deepen our understanding of this critical facet in cancer prognosis. We conducted a retrospective, population-based cohort study including 493 patients with stage II colon cancer and evaluated tumor budding using IHC, following the H&E-based guidelines proposed by the International Tumor Budding Consensus Conference Group. Correlation between H&E-based and IHC-based tumor budding was assessed using a four-tiered scoring system that included a zero budding (Bd0) category. Survival analyses explored the prognostic significance of tumor budding assessed by IHC and H&E. As expected, IHC-based tumor budding evaluation yielded significantly higher bud counts compared to H&E (p < 0.01). Interestingly, 21 patients were identified with no tumor budding using IHC. This was associated with significantly improved recurrence-free survival (HR = 5.19, p = 0.02) and overall survival (HR = 4.47, p = 0.04) in a multivariate analysis when compared to tumors with budding. The Bd0 category demonstrated a 100% predictive value for the absence of recurrence. In conclusion, IHC-based tumor budding evaluation in stage II colon cancer provides additional prognostic information. The absence of tumor budding is associated with a favorable prognosis and may serve as a potential marker for identifying patients with no risk of recurrence.

Tumor budding -a potential biomarker in low grade salivary gland carcinomas?

Low-grade salivary gland carcinoma is regularly treated with surgical therapy of the salivary gland without elective neck dissection in T1/2 carcinomas, either alone or with adjuvant radiation therapy. However, occult metastasis and locoregional recurrence influence therapy and outcome. Tumor budding is an emerging prognostic pathological factor in many carcinomas, but has not yet been adequately considered in salivary gland carcinomas. We conducted a retrospective single-center study of 64 patients diagnosed with low-grade carcinoma of the major salivary glands treated between 2003 and 2017. Pathological risk factors and TNM classification were thoroughly assessed for each case. All hematoxylin and eosin (HE)-stained histological specimens underwent careful examination, and tumor budding was identified following the guidelines set forth by the International Tumor Budding Consensus Conference in 2016. Tumor budding was not statistically significant concerning 5-year survival rate (5-YSR) (p=0.969) and mean overall survival (log-rank p=0.315). Whereas 5-year disease-free survival rate (5-YDFSR) was 87% in the low tumor budding group and 61.1% in the intermediate and high tumor budding group (p=0.021). Mean disease-free survival accounted for 100.2 months (CI: 88.6;111.9) in the low budding score group and 58.7 months (CI: 42.8;74.6) in the other group (log-rank p=0.032). Notably, pT1/2 showed significantly lower tumor buds than pT3/4 stages (2.43 tumor buds/0.785 mm2 vs. 4.19 tumor buds/0.785 mm2, p=0.034). Similar findings were noted comparing nodal-positive and nodal-negative patients, as well as patients with and without lymphovascular invasion and perineural invasion (each p<0.05). Tumor budding might be used as an additional prognostic factor for recurrence in low-grade salivary gland carcinoma, seemingly associated with a higher nodal metastasis rate and advanced tumor stages and a worse 5-YDFSR. Consequently, the evaluation of tumor budding in resection specimens of low-grade salivary gland tumor may prove valuable in decision-making for neck dissection and follow-up strategy.

A study of tumor budding and the factors, affecting interpretability of peritumoral budding, based on endoscopic colorectal biopsies from the left and right sided colorectal carcinoma

In this paper we summarize the results of a retrospective investigation of tumor budding in endoscopic preoperative biopsies in patients with colorectal adenocarcinoma (CRC). The aim of this study was to assess the peritumoral budding evaluation interpretability, based on endoscopic colorectal biopsies in left and right sided colorectal carcinoma and some of the factors that influence it. A group of 100 patients, with preoperative endoscopic biopsies was selected and the tumor buds were counted on H&amp;E stained slides, according to contemporary clinical recommendations. In the studied patient group, budding was identified in a total of 13 patients (13% of the studied cases). The primary localization of the tumor in the left or right colon was not associated with the reporting of budding in endoscopic biopsies of colorectal carcinoma. No budding was observed in highly differentiated tumors, and the presence of budding was reported less frequently in poorly differentiated tumors compared to moderately differentiated ones. Budding was accessible for evaluation in nearly 1/10 of the small biopsies from CRC. Artifacts from the sample management as well as factors related to the tumor characteristics predetermined the possibility for evaluation of tumor budding on small biopsies.

Analyzing Tumor Budding Scores in Invasive Breast Carcinoma: A Tertiary Care Center Study in South India

ABSTRACT Introduction and Aim: Tumor budding is a distinctive phenomenon which involves the presence of small clusters or individual cancer cells at the invasive front of tumors. Tumor budding has garnered attention due to its potential implications for prognosis, treatment strategies, and our understanding of cancer progression. Our aim is to study the distribution of tumor buds and its scoring in patients with infiltrating breast carcinoma and to associate with other histopathological parameters like the size of the tumor, its grade, lymphovascular invasion, and lymph node metastasis. Materials and Methods: This was a study analyzing the data of 70 resected specimens of primary breast carcinomas and providing a descriptive overview. Tumor budding was recognized, counted, and graded in hematoxylin and eosin slides. The cases were classified as low (0–4), intermediate (5–9), and high (≥10 buds) based on the count of tumor buds. Tumor budding has significant correlation with tumor grade and tumor size. Results: Of the 70 cases, 60 cases (85.71%) were diagnosed as invasive ductal carcinoma NOS. The majority [38 (54.28%)] of the cases showed an intermediate tumor budding score of 5–9/10 HPF. Conclusion: Evaluation of tumor budding allows pathologists and oncologists to gather valuable information about the tumor’s biological aggressiveness and potential for metastasis. It also helps in better risk stratification of patients, enabling a more personalized and tailored approach to treatment planning. In conclusion, assessing tumor budding in breast carcinoma holds significant clinical importance in the management and prognosis of this disease.

Preoperative Noninvasive Evaluation of Tumor Budding in Rectal Cancer Using Multiparameter MRI Radiomics

To assess the value of a multiparametric magnetic resonance imaging (MRI)-based model integrating radiomics features with clinical and MRI semantic features for preoperative evaluation of tumor budding (TB) in rectal cancer. A total of 120 patients with pathologically confirmed rectal cancer were retrospectively analyzed. The patients were randomized into training and validation cohorts in a 6:4 ratio. Radiomics features were extracted and selected from preoperative T2-weighted imaging (T2WI), diffusion-weighted imaging (DWI), and contrast-enhanced T1-weighted imaging (T1CE) sequences, after which the corresponding radiomics score (RS) was calculated, and the radiomics models (T2WI model, DWI model, and T1CE model) were constructed. Logistic regression analysis was selected to develop a combined model integrated RST2WI, RSDWI, RST1CE, and clinical and MRI semantic features. The efficacy of each model in diagnosing TB grade was observed by the receiver operating characteristic (ROC) curve. Decision curve analysis (DCA) was used to assess the clinical benefits of the models. Seven features were extracted and selected from each T2WI, DWI, and T1CE sequence to calculate the corresponding RS and construct the corresponding radiomics model. MRI reported N stagewas an independent risk factor for TB. The area under the ROC curveof the combined model was 0.961 and 0.891 in the training and validation cohorts, respectively. The combined model showed better performance than the other models. DCA showed that the net benefit of the combined model was better than that of the other models in the vast majority of threshold probabilities. A combined model integrating radiomics features and MRI semantic features allows for noninvasive preoperative evaluation of TB grading in patients with rectal cancer.

New Histopathologic Risk Model for Early T-stage Oral Squamous Cell Carcinoma: Focusing on a Modified Worst Pattern of Invasion System and a New Tumor Budding Score.

Oral squamous cell carcinoma (OSCC) is treated based on the TNM staging. However, early T-stage OSCC still exhibits substantial nodal metastasis and death rates. Recent literature highlights the independent prognostic value of worst pattern of invasion (WPOI) and tumor budding in OSCC. Nevertheless, WPOI-5 is uncommon in early T-stage OSCC, and the definitions of tumor budding and WPOI-4 overlap. Moreover, WPOI assessment is subjective, and tumor budding evaluation varies across studies. To address these limitations, we aimed to develop a modified WPOI system and a novel tumor budding scoring system that assesses single cells and high-density tumor budding. We also evaluated a new histopathologic risk model for early T-stage OSCC. The study cohort comprised 37 pT1 and 64 pT2 OSCCs. The modified WPOI demonstrated superior interobserver agreement compared with the original system (κ value: 0.98 vs. 0.53). In the multivariate analysis, modified WPOI and tumor budding score were independent prognostic factors for nodal metastasis and disease-free survival, while modified WPOI predicted disease-specific survival. By integrating these factors, our risk model stratified the patients into 3 groups. Notably, the intermediate-risk and high-risk groups exhibited significantly higher rates of nodal metastasis, recurrence, and tumor-related death. Conversely, none in the low-risk group had nodal metastasis or succumbed to the disease. Our model offered simplified scoring and potentially improved prognostic predictions. In conclusion, we've developed a modified WPOI system, a new tumor budding scoring system, and a reliable risk model that classifies early T-stage OSCC patients into distinct risk groups with significant prognostic differences.

Predictive value of tumor budding in head and neck squamous cell carcinoma: an update.

Head and neck squamous cell carcinoma forms an anatomically and functionally complex group of malignancies. The significant local aggressiveness and frequent regional relapses motivate ongoing research to identify more reliable and sensitive prognostic and predictive biomarkers. One emerging area of cancer biology is the evaluation of tumor budding at the advancing invasive front of various types of epithelial cancers. Recent studies suggest that tumor budding is a relatively common phenomenon in cancer progression and that it may have important prognostic implications for patients due to its potential to provide valuable insights into the biology and clinical behavior of head and neck cancer. In this review, we aim to provide information about tumor budding in head and neck squamous cell carcinoma. Thus, we hope to shed light on the complex biology of these malignancies, as well as aiding diagnostic, classification, and better characterization and thereby, looking for new avenues for improving patient outcomes.

Evaluation of tumor budding and its correlation with histomorphological prognostic markers in oral squamous cell carcinoma and its association with the epithelial-mesenchymal transition process.

To evaluate the association of tumor budding (TB) with prognostic histomorphological parameters in oral squamous cell carcinoma (OSCC) and to investigate the correlation of TB intensity with epithelial to mesenchymal transition (EMT). A total of 200 cases diagnosed as OSCC were selected and their TB status was reviewed using Hematoxylin and eosin (H and E) and Immunohistochemistry (IHC). Correlation with histomorphological prognostic parameters was done. Also, IHC for Vimentin and E-cadherin was performed to look for EMT. On H and E examination, TB was observed in 154/200 (77%). About 88/154 (57.14%) cases showed a high TB (>5 TB/10 hpf) which increased to 100/154 (64.9%) cases on IHC staining. The intensity of TB was significantly associated with tumor grade and depth of invasion. It was also significantly associated with reduced expression for E-Cadherin and upregulation of Vimentin establishing a pathogenetic correlation between the TB and EMT. Therefore, our results suggest that TB is associated with poor prognosis and histologically represents EMT in OSCC which further adds to the aggressiveness of the tumor.

Tumor budding as an indicator for lymph node metastasis and prognosis of early gastric cancer.

Tumor budding, considered as an independent risk factor reflecting prognosis of some malignant tumors, has been recognized as an important clinicopathological indicator of colorectal carcinoma. However, the evaluation of tumor budding and its clinicopathological significance in gastric cancer remain controversial. To investigate the relationship between tumor budding and clinical biological behavior of early gastric cancer (EGC) and assess the predictive value of tumor budding for lymph node metastasis as well as its impact on prognosis of EGC patients. Tissue specimens of 164 EGC patients who underwent radical gastrectomy between June 2011 and January 2017 from a single center were selected to carry out HE and CK staining respectively, so as to evaluate tumor budding under light microscopy. Clinicopathological data and follow-up results of all EGC patients were collected for statistical analysis among tumor budding, EGC clinicopathological factors and prognosis. Of all 164 EGC patients, there were 84 (51.2%) cases with mucosal invasion and 80 (48.8%) cases with submucosal invasion. Meanwhile, 32 cases (19.5%) had lymph node metastasis, 19 (11.6%) had lympho-vascular invasion and 4 (2.4%) had early recurrence. Tumor budding were observed in 90 (54.9%) patients, with low-grade budding 68 (41.5%) cases and high-grade budding 22 (13.4%) cases. Tumor budding was closely correlated with tumor size (c2 = 6.609, P = 0.037), tumor histologic differentiation (c2 = 10.522, P = 0.032), depth of invasion (c2 = 8.787, P = 0.012), lymph node metastasis (c2 = 24.226, P < 0.01), TNM stage (c2 = 24.226, P < 0.01), lympho-vascular invasion (c2 = 8.225, P = 0.016) and early recurrence (c2 = 6.462, P = 0.040). Additionally, tumor budding was correlated with postoperative survival rate as well. Multiple regression analysis revealed that tumor budding was an independent influencing factor of postoperative 3-year survival rate, 5-year survival rate, OS, DFS and DSS (P < 0.05). Furthermore, tumor budding was an independent risk factor of lymph node metastasis of EGC patients, and high-grade budding was a high-risk indicator of lymph node metastasis. Tumor budding is related to tumor size, tumor histologic differentiation, depth of invasion, lymph node metastasis, lympho-vascular invasion and early recurrence of EGC. Tumor budding, especially high-grade budding can serve as an indicator for predicting lymph node metastasis of EGC, and high-grade budding could be an important parameter for evaluating prognosis of EGC patients.

To investigate the prognostic factors of stage Ⅰ-Ⅱ gastric cancer based on P53 mutation and tumor budding.

P53 is a tumor suppressor and genome guardian factor, and tumor budding is a key link in tumor metastasis. The purpose of this study was to investigate P53 mutation and tumor budding in stage Ⅰ-Ⅱ gastric cancer, to explore the correlation with clinicopathological features, and to reveal the independent prognostic factors of gastric cancer. The data of 588 patients with stage Ⅰ-Ⅱ gastric cancer who underwent radical surgical resection from April 2015 to December 2016 in the Fourth Hospital of Hebei Medical University were retrospectively analyzed and followed up. Immunohistochemistry Envision method was used to conduct P53 staining for paraffin-embedded gastric cancer tissues, and ITBCC recommended tumor budding evaluation method was used to count tumor budding in gastric cancer tissues. The factors affecting the prognosis of gastric cancer were analyzed. There were 209 cases (35.54%) of P53 wild-type and 379 cases (64.46%) of P53 mutant in 588 patients with stage Ⅰ-Ⅱ gastric cancer. P53 mutation rate were closely correlated with Lauren classification (χ2 =8.152, p=0.017), degree of differentiation (χ2 =10.495, p=0.004), number of lymph node metastasis (χ2 =25.550, p<0.001), and clinical stage (χ2 =7.617, p=0.016). Tumor budding were closely correlated with Lauren classification (χ2 =194.533, p<0.001), degree of tissue differentiation (χ2 =22.719, p<0.001), depth of tumor invasion (χ2 =19.204, p=0.004), number of lymph node metastasis (χ2 =22.555, p=0.001), clinical stage (χ2 =10.769, p=0.005), and vascular tumors bolt (χ2 =12.478, p=0.002). The higher the tumor budding grade was, the higher the P53 mutation rate was (χ2 =12.933, p=0.002). Lauren classification (p<0.001), degree of tissue differentiation (p=0.005), vascular tumors bolt (p<0.001) and P53 mutation (p=0.006) were independent influencing factors for 5-year survival of patients with stage Ⅰ-Ⅱ gastric cancer. P53 mutation status is an independent prognostic factor for gastric cancer patients and a promising cancer treatment target. Tumor budding is a very reliable independent prognostic parameter with important clinical value and should be routinely reported as a biomarker.

EPDR1 levels and tumor budding predict and affect the prognosis of bladder carcinoma

BackgroundBladder carcinoma is a common malignancy of the urinary system. The previous study showed that EPDR1 expression was significantly related to the carcinogenesis and progression of bladder carcinomaMethodsWe retrospectively reviewed the records of 621 patients who were newly diagnosed with bladder carcinoma between January 2018 and August 2020 at The Affiliated Hospital of Zunyi Medical University. We conducted immunohistochemistry of EPDR1 in tumor tissues. Meanwhile, tumor budding evaluation was also carried out by 2 independent experienced pathologists.Results80 patients were included in this study with a median age of 66 years (range; 42–88 years). 45% of the patients (36/80) were non-muscle-invasive bladder carcinoma patients, while 55% of muscle-invasive bladder carcinoma(44/80). The follow-up time was from 6 months to 36 months. We found that there were significant differences in expression of EPDR1 in the tumor pT stages(p<0.05), pM stages(p<0.05), and pN stages(p<0.05). Meanwhile, a higher expression of EPDR1 indicated a worse outcome for the patient(p<0.05). A tendency toward a worse status of the patient was accompanied by a high positive rate (p<0.001). Moreover, the IOD of EPDR1 had a positive relationship with TB (p<0.05). Furthermore, we found that EPDR1 and tumor budding could be crucial factors for affecting the prognosis of bladder carcinoma, even better than pTMN(Riskscore=(0.724)* pT_stage +(4.960) *EPDR1+(4.312)*TB).ConclusionIn conclusion, bladder cancer patients with higher expression levels of EPDR1 had worse survival outcomes. The combination of TB and EPDR1 levels could predict the prognosis for muscle-invasive bladder cancer patients.

Prediction of tumor budding in patients with rectal adenocarcinoma using b-value threshold map.

To investigate the feasibility of b-value threshold (bThreshold) map in preoperative evaluation of tumor budding (TB) in patients with locally advanced rectal cancer (LARC). Patients with LARC were enrolled and underwent diffusion-weighted imaging (DWI). Contrast-to-noise ratio (CNR) between the lesions and normal tissues was assessed using DWI and bThreshold maps. TB was counted and scored using hematoxylin and eosin staining. Reproducibility for the apparent diffusion coefficient (ADC), bThreshold values, and region-of-interest (ROI) sizes were compared. Differences in ADC and bThreshold values with low-intermediate and high TB grades and the correlations between mean ADC and bThreshold values with TB categories were analyzed. Diagnostic performance of ADC and bThreshold values was assessed using area under the curve (AUC) and decision curve analysis. Fifty-one patients were evaluated. The CNR on bThreshold maps was significantly higher than that on DW images (9.807 ± 4.811 vs 7.779 ± 3.508, p = 0.005). Reproducibility was excellent for the ADC (ICC 0.933; CV 8.807%), bThreshold values (ICC 0.958; CV 7.399%), and ROI sizes (ICC 0.934; CV 8.425%). Significant negative correlations were observed between mean ADC values and TB grades and positive correlations were observed between mean bThreshold values and TB grades (p < 0.05). bThreshold maps showed better diagnostic performance than ADC maps (AUC, 0.914 vs 0.726; p = 0.048). In LARC patients, bThreshold values could distinguish different TB grades better than ADC values, and bThreshold maps may be a preoperative, non-invasive approach to evaluate TB grades. • Compared with diffusion-weighted images, bThreshold maps improved visualization and detection of rectal tumors. • Agreement and diagnostic performance of bThreshold values are superior to apparent diffusion coefficient in assessing tumor budding grades in patients with locally advanced rectal cancer. • bThreshold maps could be used to evaluate tumor budding grades non-invasively before operation.

Tumor budding may be a promising prognostic indicator in intrahepatic cholangiocarcinoma: A multicenter retrospective study.

This retrospective study evaluated our hypothesis that high tumor budding (≥10 buds) may help determine the appropriate T category for more accurate staging of intrahepatic cholangiocarcinoma (ICC). We analyzed the clinical and histopathologic data of 235 consecutive patients with histologically confirmed ICC following hepatectomy at five university hospitals in the Kansai region of Japan between January 2009 and December 2020. ICC staging was based on the Liver Cancer Study Group of Japan (LCSGJ) staging system, 6th edition. Patients with ICC with high budding showed significantly shorter disease-specific survival (DSS) and disease-free survival (DFS) than patients with low/intermediate budding. Cox proportional hazards regression analysis showed a hazard ratio of 2.2-2.3 (P < 0.05) for high budding. Based on these results, we modified the T category of ICC in the LCSGJ staging system by adding severity of tumor budding as a fourth determinant. This proposed staging system for ICC has significantly improved the prognostic accuracy for both DSS and DFS (both: P < 0.05). High tumor budding is a new candidate for an additional determinant of the T category in staging ICC. An LCSGJ staging system containing an additional evaluation of tumor budding may lead to improved staging accuracy.

The feasibility of MRI-based radiomics model in presurgical evaluation of tumor budding in locally advanced rectal cancer.

To build and validate a magnetic resonance imaging-based radiomics model to preoperatively evaluate tumor budding (TB)in locally advanced rectal cancer(LARC). Pathologically confirmed LARC cases submitted to preoperative rectal MRI in two distinct hospitals were enrolled in this retrospective study and assigned to cohort 1 (training set, n = 77; test set, n = 51) and cohort 2 (validation set, n = 96). Radiomics features were obtained from multiple sequences, comprising high-resolution T2, contrast-enhanced T1, and diffusion-weighted imaging (T2WI, CE-T1WI, and DWI, respectively). The least absolute shrinkage and selection operator (LASSO)was utilized to select the optimal features from T2WI, CE-T1WI, DWI, and the combination of multi-sequences, respectively. A support vector machine (SVM)classifier was utilized to construct various radiomics models for discriminating the TB grades. Receiver operating characteristic curve analysis and decision curve analysis (DCA)were carried out to determine the diagnostic value. Five optimal features associated with TB grade were determined from combined multi-sequence data. Accordingly, a radiomics model based on combined multi-sequences had an area under the curve of 0.796, with an accuracy of 81.2% in the validation set, showing a better performance in comparison with other models in both cohorts (p < 0.05). DCA exhibited a clinical benefit for this radiomics model. The novel MRI-based radiomics model combining multiple sequences is an effective and non-invasive approach for evaluating TB grade preoperatively in patients with LARC.

Immunohistochemical Expression of “HCG-β” in Colorectal Carcinoma

BACKGROUND: Tumor budding is associated with adverse histology and is a predictor of lymph node metastasis. Human chorionic gonadotropin-beta (hCG-β) expression in non-trophoblastic tumors has been associated with aggressive behavior. AIM: Evaluation of tumor budding and hCG-β _immunohistochemical expression in colorectal carcinoma (CRC), and correlation of their expression with various clinicopathological parameters. MATERIALS AND METHODS: Immunohistochemical staining for hCG-β _was performed on paraffin-embedded sections of 60 cases of CRC. Tumors with cytoplasmic or membranous staining of more than five epithelial cell clusters were designated hCG-β _positive; otherwise, they were designated hCG-β _negative. Tumor budding was assessed in hematoxylin and eosin stained slides and was classified as; low: 0–4 buds, intermediate: 5–9 buds and high: ≥10 buds; with exclusion of pure mucoid or signet ring cell morphology cases from analysis. RESULTS: Tumor budding was low in (58.8%) of the cases, intermediate in (15.7%), and high in (25.5%). There was a statistically significant correlation between tumor budding and tumor histological grade (p = 0.011), lymph node metastasis (N) (p = 0.009), overall pathologic stage group (p = 0.009), modified Dukes’ stage (p = 0.009), lymphovascular invasion (p = 0.000), and desmoplastic reaction (p = 0.004). Positive hCG-β _alpha expression was detected in 12 (20%) of cases. There were statistically significant correlations between hCG-β _expression and each of lymphovascular invasion (p = 0.042) and tumor budding (p = 0.000). CONCLUSION: hCG-β _is a marker of aggressiveness that may have essential role in tumor invasion. Tumor budding is crucial event in tumor invasion and metastasis. Tumor budding with hCG-β _expression is a novel prognostic parameter and may represent a potential therapeutic target.

Prognostic significance of tumor budding in muscle invasive urothelial carcinomas of the urinary bladder

ObjectiveThe aim of the present study was to analyze the prognostic significance of tumor budding in muscle-invasive urothelial carcinomas of the urinary bladder, and also to determine an optimal threshold value in evaluation. Patients and methodsThe study included 108 patients diagnosed with muscleinvasive conventional urothelial carcinoma between 2010 and 2020. Tumor budding was evaluated on H&E-stained slides. The critical tumor budding number was determined with the “receiver operating characteristics (ROC)” curve. Cases with a tumor budding number of ≤6 were categorized as low, and cases with >6 as high tumor budding. ResultsThe univariate Cox proportional hazards regression model for recurrence-free survival showed that lymphovascular invasion (P = 0.001), tumor budding (P = 0.012), pT stage (T4 vs. T2) (P = 0.005), and lymph node metastasis (P = 0.009) were significantly associated with recurrence-free survival. The multivariate Cox proportional hazards regression model utilizing backward stepwise (wald) method revealed that only LVI (P = 0.001) was independent risk factor for recurrence-free survival. The univariate Cox analysis showed that lymphovascular invasion (P = 0.001), tumor budding (P = 0.004), pT stage (T4 vs. T2) (P = 0.003), and lymph node metastasis (P = 0.001) were significantly associated with overall survival. The multivariate Cox analysis (backward stepwise (wald) method) revealed that tumor focality (P = 0.018), pT stage (T4 vs. T2) (P = 0.015), and lymphovascular invasion (P = 0.002) were independent factors for overall survival. ConclusionsOur findings suggested that the evaluation of tumor budding may be a useful parameter for predicting outcome in patients with muscle-invasive bladder cancer.

Association of Tumor Budding With Immune Evasion Pathways in Primary Colorectal Cancer and Patient-Derived Xenografts

Tumor budding has been found to be of prognostic significance for several cancers, including colorectal cancer (CRC). Additionally, the molecular classification of CRC has led to the identification of different immune microenvironments linked to distinct prognosis and therapeutic response. However, the association between tumor budding and the different molecular subtypes of CRC and distinct immune profiles have not been fully elucidated. This study focused, firstly, on the validation of derived xenograft models (PDXs) for the evaluation of tumor budding and their human counterparts and, secondly, on the association between tumor budding and the immune tumor microenvironment by the analysis of gene expression signatures of immune checkpoints, Toll-like receptors (TLRs), and chemokine families. Clinical CRC samples with different grades of tumor budding and their corresponding PDXs were included in this study. Tumor budding grade was reliably reproduced in early passages of PDXs, and high-grade tumor budding was intimately related with a poor-prognosis CMS4 mesenchymal subtype. In addition, an upregulation of negative regulatory immune checkpoints (PDL1, TIM-3, NOX2, and IDO1), TLRs (TLR1, TLR3, TLR4, and TLR6), and chemokine receptors and ligands (CXCR2, CXCR4, CXCL1, CXCL2, CXCL6, and CXCL9) was detected in high-grade tumor budding in both human samples and their corresponding xenografts. Our data support a close link between high-grade tumor budding in CRC and a distinctive immune-suppressive microenvironment promoting tumor invasion, which may have a determinant role in the poor prognosis of the CMS4 mesenchymal subtype. In addition, our study demonstrates that PDX models may constitute a robust preclinical platform for the development of novel therapies directed against tumor budding in CRC.

The prognostic value of tumor budding in laryngeal squamous cell carcinoma.

BackgroundThe predictive value of tumor budding in several cancers is of the essence. The 5-year survival of laryngeal squamous cell carcinoma patients is decreasing despite the improvement of therapy. In order to help improve the prognosis of LSCC patients, we aimed to investigate the value of tumor budding on the prognosis in laryngeal squamous cell cancer (LSCC) by the pathological characteristics of the surgical cases.MethodsArchival clinical specimens of 51 patients diagnosed with LSCC were included in the research. On the basis of hematoxylin and eosin (H&E)-stained slides, tumor-stroma ratio (TSR), tumor budding and nuclear features were assessed. Correlation between clinical data and histologic characteristics was carried out using the Kaplan-Meier method and the Cox proportional hazards regression model, respectively.ResultsTotal tumor budding was independent prognostic parameter of 5-year recurrence-free survival (RFS) and overall survival (OS) in LSCC. The evaluation of tumor budding can be as a part of the routine histopathological report in LSCC.ConclusionsTumor budding can be an independent factor of prognosis of LSCC patients and should be as a part of routine histopathologic report for LSCC cases.

High-grade intratumoral tumor budding is a predictor for lymphovascular invasion and adverse outcome in stage II colorectal cancer.

Evaluation of tumor budding in colorectal cancer (CRC) may help to predict the tumors' metastatic potential and patients with an aggressive tumor, although not yet metastasized at time of surgery might benefit from adjuvant therapy. The degree of intratumoral tumor budding (ITB) was classified as low, intermediate, and high grade according to the recommendations of the International Tumor Budding Consensus Conference (ITBCC) 2016 on H&E and pankeratin-stained TMA sections from 1262 CRC, no special type (NST), including 655 stage II CRC and was correlated to clinicopathological data and overall survival. Results show that higher ITB rates are significantly linked to higher tumor grade and stage, positive nodal status, lymphovascular invasion (P< 0.0001 each), absence of peritumoral lymphocytes, infiltrating type invasive tumor margin, left-sided cancer localization, and mismatch-repair proficient cancers (P<0.05 each). In a cohort of 655 stage II CRC, ITB was associated with lymphovascular invasion (P= 0.0459) and adverse clinical outcome (P< 0.0001). In a multivariate analysis including tumor stage, tumor grade, lymphovascular invasion, ITB, and tumor localization, only low tumor stage (P= 0.0022) and absence of lymphovascular invasion (P= 0.0043) showed independent prognostic significance. In conclusion, our findings argue towards a clinical utility of ITB as a prognostic biomarker in stage II colorectal cancer to define patients who might benefit from adjuvant therapy. ITB might be used as additional or surrogate marker in CRC in which peritumoral tumor budding is difficult to assess.