Desmoplastic Stromal Reaction Research Articles

Primary Tumor Histology is a Potential Independent Predictor of Oncological outcomes in Colorectal Cancer

Abstract Introduction/Objective Evaluation of the primary tumor histology can identify features in the tumor microenvironment (TME) that are intricately linked to molecular changes and aggressive tumor behavior in colorectal cancer (CRC). In this study, we evaluated the primary tumor histology such as the tumor histologic subtype, the presence of poorly differentiated clusters (PDCs), and stromal desmoplastic reaction (SDR), and determined their relationship with tumor grade, stage, and DNA mismatch repair (DNA MMR) gene status. Methods/Case Report This was a retrospective study. Hematoxylin and eosin-stained microarray sections of CRC tissues in 208 patients were pathologically evaluated. PDC was defined as a cluster of ≥ 5 nuclei in the stroma with no glandular differentiation. SDR was defined as stromal connective tissue remodeling. Parallel sections were stained with antibodies against the MLH1, PMS2, HMS6, and HMS2 for their expression. Statistical analysis was performed for the association between the variables Results (if a Case Study enter NA) There were 208 patient samples, 113 were males (54.3%) and 95 were females (45.7%). 61.5% were ≥ 50 years and 38.5% were less than 50 years. 59.4% and 37.5% of stages II and III tumors had PDCs. Univariate analysis showed a strong association between tumor grade, stage, and SDR (p=0.005 and p= 0.013 respectively) but not with PDC and MMR deficiency. 16.8% of tumors had MMR deficiency which was more prevalent in adenocarcinoma with mucinous components (27%). Multivariate analysis showed a strong association with PDC and tumor stage (p=0.026), tumor subtype and stage and grade (p=0.007, p= 0,023), stage, and SDR (p = 0.026). Grade, stage, SDR, and MMR all showed very strong association (p=0.005, p=0.019). Conclusion Our data shows that tumor histology is an independent predictor of oncological outcomes in CRC. Future longitudinal studies on prospective cohorts are required to validate our observations and increase the power of our deductions.

The Concept of Stroma AReactive Invasion Front Areas (SARIFA) as a new prognostic biomarker for lipid-driven cancers holds true in pancreatic ductal adenocarcinoma

BackgroundPancreatic ductal adenocarcinoma (PDAC) is a ‘difficult-to-treat’ entity. To forecast its prognosis, we introduced a new biomarker, SARIFA (stroma areactive invasion front areas), which are areas at the tumour invasion front lacking desmoplastic stroma reaction upon malignant invasion in the surrounding tissue, leading to direct contact between tumour cells and adipocytes. SARIFA showed its significance in gastric and colorectal carcinoma, revealing lipid metabolism alternations that promote tumour progression.MethodsWe reviewed the SARIFA status of 166 PDAC cases on all available H&E-stained tumour slides from archival Whipple-resection specimens. SARIFA positivity was defined as SARIFA detection in at least 66% of the available slides. To investigate alterations in tumour metabolism and microenvironment, we performed immunohistochemical staining for FABP4, CD36 and CD68. To verify and quantify a supposed delipidation of adipocytes, adipose tissue was digitally morphometrised.ResultsIn total, 53 cases (32%) were classified as SARIFA positive and 113 (68%) as SARIFA negative. Patients with SARIFA-positive PDAC showed a significantly worse overall survival compared with SARIFA-negative cases (median overall survival: 11.0 months vs. 22.0 months, HR: 1.570 (1.082–2.278), 95% CI, p = 0.018), which was independent from other prognostic markers (p = 0.014). At the invasion front of SARIFA-positive PDAC, we observed significantly higher expression of FABP4 (p < 0.0001) and higher concentrations of CD68+ macrophages (p = 0.031) related to a higher risk of tumour progression. CD36 staining showed no significant expression differences. The adipocyte areas at the invasion front were significantly smaller, with mean values of 4021 ± 1058 µm2 and 1812 ± 1008 µm2 for the SARIFA-negative and -positive cases, respectively (p < 0.001).ConclusionsSARIFA is a promising prognostic biomarker for PDAC. Its assessment is characterised by simplicity and low effort. The mechanisms behind SARIFA suggest a tumour-promoting increased lipid metabolism and altered immune background, both showing new therapeutic avenues.

Correlation analysis of interstitial maturity and prognosis of colorectal cancer: Meta-analysis.

To investigate the relationship between interstitial maturity and prognosis of colorectal cancer. To examine the correlation between interstitial maturity and the prognosis of colorectal cancer. The paper database PubMed, EMBASE, Cochranelibrary, Springerlink, CNKI, and Wanfang database were searched until December 2023. "tumor stroma maturity" "desmoplastic stroma reaction" "desmoplastic reaction" "stroma reaction" "degree of stroma reaction "" stroma classification" "stroma density" "colorectal cancer" "colon cancer" "rectal cancer" "prognosis" were searched for the search terms. Two system assessors independently screened the literature quality according to the inclusion exclusion criteria, Quality evaluation and data extraction were performed for the included literatures, and meta-analysis was performed for randomized control trials included at using Review Manager 5.2 software. Finally, data of 9849 patients with colorectal cancer from 19 cosets in 15 literatures were included, including 4339 patients with mature type (control group), 3048 patients with intermediate type (intermediate group) and 2456 patients with immature type (immature group). The results of meta-analysis showed: Relapse-free survival [hazard ratio (HR) = 2.66, 95% confidence interval (CI): 2.30-3.08; P < 0.00001], disease-free survival (HR = 3.68, 95%CI: 2.33-5.81; P < 0.00001) and overall survival (HR = 1.70, 95%CI: 1.53-1.87; P < 0.00001) were significantly lower than those in mature group (control group); relapse-free survival (HR = 1.36, 95%CI: 1.17-1.59; P < 0.0001) and disease-free survival rate (HR = 1.85, 95%CI: 1.53-2.24; P < 0.0001) was significantly lower than the mature group (control group). There is the correlation between tumor interstitial maturity and survival prognosis of colorectal cancer, and different degrees of tumor interstitial maturity have a certain impact on the quality of life of colorectal cancer patients.

Evaluation of the significance of tumor stromal patterns and peri-tumoral inflammation in head and neck squamous cell carcinoma with special reference to the Yamamoto-Kohama classification.

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide with 878,348 new cases. Cancer-associated fibroblasts (CAFs) are the predominant cell type in tumor stroma and are important promoters of tumor progression. The aim of the study was to evaluate the pattern of desmoplastic stromal reaction and peri-tumoral inflammatory infiltrate with the histological grade and clinical data. A total of 60 cases of HNSCC were included in the study. The hematoxylin and eosin (H and E)-stained sections from all cases were examined by two experienced pathologists for the grade, nature of stomal reaction (SR), peri-tumoral inflammatory infiltration, Yamamoto-Kohama classification grade, worst pattern of invasion (WPOI), depth of invasion (DOI), and other histopathological parameters. Correlation analysis was conducted using the Chi-square test. P- value less than 0.05 was considered statistically significant. Immature SR was not observed in any of the well-differentiated squamous cell carcinoma (SCC) cases. However, one (3.7%) case of moderately differentiated SCC and two (28.6%) cases of poorly differentiated SCC showed signs of immature SR. In the case of the higher grades of the YK classification, specifically grades 4C and 4D, a more profound depth of tumor cell invasion, equal to or exceeding 10 mm, was evident in six (66.67%) and two (28.57%) cases, respectively. Additionally, among the seven (11.7%) cases classified as poorly differentiated carcinoma, three (42.85%) displayed a WPOI score of 5. SR and the tumor invasive pattern in HNSCC are related to prognosis and may indicate tumor aggressiveness.

Histological parameters and stromal desmoplastic status affecting accurate diagnosis of extraprostatic extension of prostate cancer using multi-parametric magnetic resonance imaging.

To investigate the clinicopathological factors affecting discrepancies between multi-parametric magnetic resonance imaging (mpMRI) and histopathological evaluation for diagnosis of extraprostatic extension (EPE) of prostate cancer. One hundred-and-three lesions from 96 cases with suspected EPE on preoperative mpMRI, of which 60 and 43 showed bulging and frank capsular breach, respectively, were grouped according to pathological (p)EPE in radical prostatectomy specimens. Additionally, clinicopathological/immunohistochemical findings for periostin reflecting a desmoplastic stromal reaction were compared between these groups. pEPE was detected in 49 (48%) of the 103 lesions. Of these, 25 (42%) showed bulging and 24 (56%) showed frank capsular breach on MRI. In the total cohort, the absence of pEPE was significantly associated with a lower Gleason Grade Group (GG) (p < 0.0001), anterior location (p = 0.003), absence of intraductal carcinoma of the prostate (IDC-P) (p = 0.026), and high stromal periostin expression (p < 0.0001). These trends were preserved in subgroups defined by MRI findings, except for anterior location/IDC-P in the bulging subgroup. GG, anterior location, and periostin expression may cause mpMRI-pathological discrepancies regarding EPE. Periostin expression was a significant pEPE-negative factor in all subgroup analyses. Our results indicate that patients with suspected EPE on MRI, regardless of their pEPE results, should be followed as carefully as those with definite pEPE.

Stroma AReactive Invasion Front Areas (SARIFA) improves prognostic risk stratification of perioperative chemotherapy treated oesophagogastric cancer patients from the MAGIC and the ST03 trial.

Tumour-associated fat cells without desmoplastic stroma reaction at the invasion front (Stroma AReactive Invasion Front Areas (SARIFA)) is a prognostic biomarker in gastric and colon cancer. The clinical utility of theSARIFA status in oesophagogastric cancer patients treated with perioperative chemotherapy is currently unknown. TheSARIFA status was determined in tissue sections from patients recruited into the MAGIC (n = 292) or ST03 (n = 693) trials treated with surgery alone (S, MAGIC) or perioperative chemotherapy (MAGIC, ST03). The relationship between SARIFA status, clinicopathological factors, overall survival (OS) and treatment was analysed. TheSARIFA status was positive in 42% MAGIC trial S patients, 28% MAGIC and 48% ST03 patients after pre-operative chemotherapy. SARIFA status was related to OS in MAGIC trial S patients and was an independent prognostic biomarker in ST03 trial patients (HR 1.974, 95% CI 1.555-2.507, p < 0.001). ST03 patients with lymph node metastasis (ypN + ) and SARIFA-positive tumours had poorer OS than patients with ypN+ and SARIFA-negative tumours (plogrank < 0.001). TheSARIFA status has clinical utility as prognostic biomarker in oesophagogastric cancer patients irrespective of treatment modality. Whilst underlying biological mechanisms warrant further investigation, theSARIFA status might be used to identify new drug targets, potentially enabling repurposing of existing drugs targeting lipid metabolism.

Glutamine addiction and therapeutic strategies in pancreatic cancer

Pancreatic cancer remains one of the most lethal diseases worldwide owing to its late diagnosis, early metastasis, and poor prognosis. Because current therapeutic options are limited, there is an urgent need to investigate novel targeted treatment strategies. Pancreatic cancer faces significant metabolic challenges, principally hypoxia and nutrient deprivation, due to specific microenvironmental constraints, including an extensive desmoplastic stromal reaction. Pancreatic cancer cells have been shown to rewire their metabolism and energy production networks to support rapid survival and proliferation. Increased glucose uptake and glycolytic pathway activity during this process have been extensively described. However, growing evidence suggests that pancreatic cancer cells are glutamine addicted. As a nitrogen source, glutamine directly (or indirectly via glutamate conversion) contributes to many anabolic processes in pancreatic cancer, including amino acids, nucleobases, and hexosamine biosynthesis. It also plays an important role in redox homeostasis, and when converted to α-ketoglutarate, glutamine serves as an energy and anaplerotic carbon source, replenishing the tricarboxylic acid cycle intermediates. The present study aims to provide a comprehensive overview of glutamine metabolic reprogramming in pancreatic cancer, focusing on potential therapeutic approaches targeting glutamine metabolism in pancreatic cancer.

Cancer-Associated Fibroblasts Influence Survival in Pleural Mesothelioma: Digital Gene Expression Analysis and Supervised Machine Learning Model.

The exact mechanism of desmoplastic stromal reaction (DSR) formation is still unclear. The interaction between cancer cells and cancer-associated fibroblasts (CAFs) has an important role in tumor progression, while stromal changes are a poor prognostic factor in pleural mesothelioma (PM). We aimed to assess the impact of CAFs paracrine signaling within the tumor microenvironment and the DSR presence on survival, in a cohort of 77 PM patients. DSR formation was evaluated morphologically and by immunohistochemistry for Fibroblast activation protein alpha (FAP). Digital gene expression was analyzed using a custom-designed CodeSet (NanoString). Decision-tree-based analysis using the "conditional inference tree" (CIT) machine learning algorithm was performed on the obtained results. A significant association between FAP gene expression levels and the appearance of DSR was found (p = 0.025). DSR-high samples demonstrated a statistically significant prolonged median survival time. The elevated expression of MYT1, KDR, PIK3R1, PIK3R4, and SOS1 was associated with shortened OS, whereas the upregulation of VEGFC, FAP, and CDK4 was associated with prolonged OS. CIT revealed a three-tier system based on FAP, NF1, and RPTOR expressions. We could outline the prognostic value of CAFs-induced PI3K signaling pathway activation together with FAP-dependent CDK4 mediated cell cycle progression in PM, where prognostic and predictive biomarkers are urgently needed to introduce new therapeutic strategies.

Low-grade Hidradenocarcinomas: A Clinicopathologic Study of an Unusual Carcinoma That Can Mimic its Benign Counterpart.

Hidradenocarcinomas are rare cutaneous adnexal malignancies with sweat gland differentiation that can show a broad spectrum of histomorphologic appearances, ranging from low to high grade. The diagnosis of low-grade hidradenocarcinoma can be challenging and may be mistaken for benign hidradenomas, especially on superficial and partial samples. We performed a retrospective analysis of 16 low-grade hidradenocarcinomas, obtained from 4 large academic institutions. All neoplasms presented clinically as nodular lesions that ranged in size from 1.5 to 6.0cm. All patients were adults and their age ranged from 33 to 74 years of age. All cases shared features similar to hidradenomas in the surface and mid portion of the tumors and all tumors had 1 or more histomorphologic clues to malignancy, including the presence of an asymmetric and infiltrative growth pattern (especially at the base of the tumors), perineurial invasion, and a desmoplastic stromal reaction. In the tumors evaluated for immunohistochemistry, the tumor cells were positive for p63, EMA, AE1/AE3, MNF116, and CK7. Three patients underwent sentinel lymph node biopsy, and 2 cases showed metastatic disease to regional lymph nodes. All cases (including the 2 cases that had regional lymph node metastasis), showed no local recurrence or distant metastasis observed after a complete re-excision of the tumors (follow-up range from 6 to 72mo). Our study highlights the salient clinical and histopathologic features of low-grade hidradenocarcinomas and emphasizes the potential diagnostic pitfalls in distinguishing this entity from other neoplasms. Our results indicate that a combination of thorough histopathologic inspection is necessary to support the diagnosis of this rare neoplasm. These tumors can be exceedingly difficult to diagnose and awareness of the subtle features of low-grade hidradenocarcinoma is of importance are as it remains a diagnostic challenge for practicing pathologists.

A-257 OMISSION OF LYMPH NODE DISSECTION IN MEDULLARY THYROID CANCER WITHOUT A DESMOPLASTIC STROMAL REACTION, COULD BE AN OPTION?

Abstract Background Predictive criteria to determine the absence of node metastases from thyroid specimens are scarce for sporadic medullary thyroid cancer (MTC). It has been suggested that lymph node dissection could be avoided in patients with MTC without a desmoplastic stromal reaction (DSR-negative). The aim of our study was to perform analyse the correlation between DSR status and lymph node involvement in order to establish a criterion on which to base prophylactic surgery in these patients. Methods This was a multicentric retrospective cohort study of a prospectively maintained database of patients with medullary thyroid cancer treated in two tertiary hospitals using a standardised protocol and subdivided into DSR-negative and -positive groups. Intraoperative, long-term clinical and biochemical follow-up data were collected, and baseline parameters and histopathological characteristics were compared between groups. Results The study included 29 patients. In the DSR-negative group (24.1% of all tumours) no patient had central or lateral lymph node or distant metastases at diagnosis or during follow-up, and all patients were biochemically cured. In the DSR-positive group (75.9% all tumours), lymph nodes were present in 66% of patients. DSR-negative tumours were more often stage pT1a and levels of basal calcitonin were significantly lower. Conclusion Lymph node surgery may be individualised in medullary thyroid cancer based on intraoperative analysis of the DSR. Patients with DSR-negative tumours may not require lymph node dissection.

Histopathological categorization of desmoplastic reaction in gallbladder carcinoma: its relation to cancer invasiveness and prognostic utility

BackgroundDesmoplastic changes in a tumor have been observed to affect the tumor microenvironment, leading to both the facilitation and prevention of tumor invasiveness.MethodsBetween 2016 and 2020, a total of 120 patients were diagnosed with gallbladder carcinoma. The cases were reviewed for the type of desmoplastic reaction, tumor grade, and stage. The type of desmoplastic reaction was classified as mature, intermediate, and immature, and they were correlated with tumor grade and stage.ResultsOut of the total number of cases, 21.67%, 48.33%, 25%, and 5% belonged to stages 1, 2, 3, and 4, respectively. A total of 79.2% of these cases showed desmoplastic stromal reaction, and there was a significant correlation (p < 0.05) between the type of desmoplasia and tumor grade and stage. The occurrence of immature stroma was correlated with increasing tumor grade and stage, and occurrence of mature stroma was negatively correlated with increasing tumor grade and stage.ConclusionThe assessment of the type of stromal desmoplastic reaction in primary gallbladder carcinoma can predict the tumor invasiveness.

Abstract 5447: Artificial intelligence (AI)-based classification of stromal subtypes reveals associations between stromal composition and prognosis in NSCLC

Abstract Background: Many cancers are known to involve a desmoplastic stromal reaction. While cancer-associated stroma (CAS) has long been appreciated histologically, single-cell molecular analyses have revealed its heterogeneity, and the cellular composition of CAS has been linked to prognosis in several cancer types, including NSCLC. While pathologists have sought to manually classify CAS based on collagen architecture or nuclear density, this approach has not sufficiently captured the heterogeneity of CAS. To this end, we have developed an artificial intelligence (AI)-based model to predict stromal composition in NSCLC from hematoxylin and eosin (H&amp;E)-stained tissue Methods: We developed a convolutional neural network-based model to classify CAS as immature, mature, densely inflamed, densely fibroblastic, or elastosis. This model was trained using manual pathologist-derived annotations (N=3019) of H&amp;E-stained whole slide images (WSIs) of PDAC obtained from the TCGA (N=126). This stromal subdivision model was deployed on H&amp;E-stained LUAD (N=468) and LUSC (N=430) WSIs. Model performance was assessed by qualitative review by expert pathologists. Human interpretable features (HIFs) were extracted from the stromal subdivision model (e.g., proportional area of mature relative to total stroma) and were assessed to identify associations with overall survival (OS) using univariate Cox regression analysis after adjusting for age, sex, and tumor stage. Results: The stromal subdivision model successfully predicted areas of immature, mature, densely inflammatory, and densely fibrotic stroma, as well as elastosis, in LUAD and LUSC. In LUAD, higher combined proportional areas of mature and fibroblastic stroma relative to total cancer stroma was associated with poor OS (p=0.007), while higher combined proportional areas of densely inflamed stroma and elastosis relative to total cancer stroma was associated with improved OS (p=0.007). These findings were validated by stratified tertile analysis based on the corresponding risk direction. Notably, while the average stromal compositions did not differ significantly between NSCLC subtypes, the stromal HIFs were only prognostic in LUAD but not in LUSC. Conclusions: We developed a first of its kind model to characterize CAS subtypes in NSCLC tissue. Features extracted from this model are related to prognosis in LUAD, but not in LUSC, further confirming the importance of CAS to tumor biology and the importance of considering histologic subtypes. Work is ongoing to identify relationships between stromal HIFs and treatment response in NSCLC as well as other cancer indications. Citation Format: Fedaa Najdawi, Sandhya Srinivasan, Neel Patel, Michael G. Drage, Christian Kirkup, Chintan Parmar, Jacqueline Brosnan-Cashman, Michael Montalto, Andrew H. Beck, Archit Khosla, Ilan Wapinski, Ben Glass, Murray Resnick, Matthew Bronnimann. Artificial intelligence (AI)-based classification of stromal subtypes reveals associations between stromal composition and prognosis in NSCLC. [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 5447.

The Token Transition-Minimal Deviation AdenocarcinomaUterine Cervix

Gastric type adenocarcinoma is a high grade, frequently discerned mucinous adenocarcinoma with gastric differentiation incriminating the endocervical canal. Of obscure aetiology, neoplasm depicts STK11 genetic mutations with chromosome 3q gain and chromosome 1p loss. Tumefaction exhibits cytological and histological features of gastric differentiation as tumour cells incorporated with voluminous, clear or pale, foamy, eosinophilic cytoplasm, tall, abundant, apical mucin, miniature, spherical, elliptical or enlarged, basal nuclei, irregular, vesicular chromatin and prominent nucleoli. The well differentiated neoplasm is comprised of well defined glandular articulations and minimal to absent stromal desmoplastic reaction. Neoplastic glands of variable outline, magnitude and irregular or claw-like glandular contour are observed. Tumour cells appear immune reactive to HIK1083, MUC6, p53, carcinoembryonic antigen (CEA) or mCEA, CK7, PAX8, CAIX or HNF-1B. Minimal deviation adenocarcinoma requires segregation from neoplasms such as human papillomavirus (HPV) associated endocervical adenocarcinoma, human papillomavirus (HPV) associated endocervical adenocarcinomas intestinal(goblet cell) type, clear cell adenocarcinoma, invasive stratified mucin producing adenocarcinoma, metastatic gastric or pancreaticobiliary carcinoma, lobular endocervical glandular hyperplasia, atypical lobular endocervical glandular hyperplasia, diffuse laminar endocervical glandular hyperplasia, tunnel clusters, florid deep glands, microglandular hyperplasia, pseudo-infiltrative tubal metaplasia, endocervicosis, endosalpingosis or endocervical type adenomyoma. Minimal deviation adenocarcinoma can be adequately categorized upon histological examination of tissue obtained with surgical sampling. Stage I tumours may be managed with surgical procedures as trachelectomy or radical hysterectomy and regional lymphadenectomy upon initial disease representation

Nested Subtype of Urothelial Carcinoma in the Upper Urinary Tract: A Series of 15 Cases.

Nested subtype of urothelial carcinoma (NVUC) of the upper urinary tract is rare. While this subtype has an aggressive behavior in the urinary bladder, little is known about this subtype in the renal pelvis and ureter. In addition, due to NVUC's bland morphology and resemblance to von Brunn nests proliferation, which can be florid in the renal pelvis and ureter, establishing a diagnosis of NVUC in these sites can be difficult, especially on small biopsies. Nineteen cases of NVUC of the renal pelvis and the ureter were identified. One was in-house and 18 cases were sent to the genitourinary pathology consult service. Four cases were excluded due to difficulty in obtaining the slides from other institutions. In all, 8/15 (53.33%) cases showed large nested morphology, 4/15 (26.66%) cases showed mixed large and small nested morphology, and only 3/15 (20%) cases showed predominantly small nested morphology. In all cases, the invasive component was pure nested subtype, and tumors were all unifocal. In 8/15 cases, there was the presence of an overlying low-grade papillary urothelial carcinoma. An associated desmoplastic stromal reaction was none/minimal in 12/15 (80%) cases, with a prominent reaction in the remaining 3/15 (20%) cases. Pathologic stages for the resections were pT1 (n=1), pT2 (n=3), pT3 (n=8), and pT4 (n=1). There was only 1 case with a lymph node metastasis at the time of resection who had pT4 disease. Follow-up was available in 10/15 cases, 7/10 cases showed no recurrences or metastases, and the remaining 3/10 cases later developed urothelial carcinoma of the bladder. In 5/15 cases, follow-up could not be obtained. Similar to NVUC in the urinary bladder, NVUC in the upper tract typically lacks an associated desmoplastic reaction and has by definition bland cytology. In contrast to NVUC of the urinary bladder, NVUC in the upper urothelial tract is: (1) more frequently associated with an overlying papillary urothelial carcinoma; (2) more commonly has a large nested pattern; and (3) appears to behave less aggressively.

Prognostic value of desmoplastic stromal reaction, tumor budding and tumor-stroma ratio in stage II colorectal cancer

Existing high-risk factors are insufficient to accurately predict the postoperative recurrence risk of stage II colorectal cancer (CRC). The discovery of additional prognostic markers may be the key to improving the current status of stage II CRC treatment. The present study aimed to evaluate the relationship among desmoplastic reaction (DR), tumor budding (TBd), the tumor-stroma ratio (TSR) and their prognostic value for relapse-free survival (RFS). In this study, 207 patients with histologically confirmed stage II CRC from January 2012 to August 2018 were retrospectively reviewed from a single center; the cohort was divided into subgroups based on low or high TSR, and low, intermediate or high DR and TBd. Kaplan-Meier curve analysis and log-rank test were applied to examine RFS among subgroups. Univariate and multivariate Cox proportional hazards analyses were used to identify independent factors associated with RFS, and a nomogram was subsequently developed. Abnormal CA242, CEA, T4 stage, presence of hypertension, internal obstruction or perforation (IOP), lymphovascular or/and perineural invasion (PNI), number of nodes examined less than 12, low-frequency microsatellite instability (MSI-L), higher Ki-67 and immature DR were associated with a lower RFS. In multivariable analysis, DR (HR =2.111; 95% CI: 1.184-3.766; P=0.011), LVI (HR =1.919; 95% CI: 1.004-3.669; P=0.049) and PNI (HR =2.724; 95% CI: 1.362-5.448; P=0.005) were prognostic factors for RFS. On this basis, a nomogram that integrated DR and clinicopathologic predictors for predicting RFS passed the calibration and had an area under the curve of 0.826. The prognostic significance of DR outperformed TBd and TSR, therefore, we recommend adding DR as a biomarker in routine pathological reports. The novel nomogram combining these factors may be used as a reliable and effective tool for the prediction of RFS in stage II CRC, thus helping optimize therapeutic regimens under cooperation of oncologists and surgeons. Further multicentric studies are required for validation of this novel, simple and cost-effective prognostic model.

Thyroid Cancer and Fibroblasts.

Simple SummaryThyroid cancer is the most common solid tumor of the endocrine glands. The cancer cell contribution to thyroid cancer development and progression has been studied extensively, whereas the involvement of the tumor microenvironment, particularly of cancer-associated fibroblasts (CAFs), in thyroid cancer growth still must be largely analyzed. The tumor microenvironment, comprising molecules, blood and lymphatic tumor vessels and several non cancer stromal cells, such as CAFs, dramatically influences solid tumor growth and therapy resistance. In particular, investigations on CAF contribution to solid tumor growth and therapeutic resistance represent an important area of oncological research. Moreover, studies focused on the role of CAFs in thyroid cancer could lead to a better comprehension of mechanisms regulating cancer growth and to the development of new therapeutic strategies. Therefore, in this paper, we review the hallmarks of CAFs and their role on thyroid cancer.Thyroid cancer is the most common type of endocrine cancer, and its prevalence continue to rise. Non-metastatic thyroid cancer patients are successfully treated. However, looking for new therapeutic strategies is of great importance for metastatic thyroid cancers that still lead to death. With respect to this, the tumor microenvironment (TME), which plays a key role in tumor progression, should be considered as a new promising therapeutic target to hamper thyroid cancer progression. Indeed, thyroid tumors consist of cancer cells and a heterogeneous and ever-changing niche, represented by the TME, which contributes to establishing most of the features of cancer cells. The TME consists of extracellular matrix (ECM) molecules, soluble factors, metabolites, blood and lymphatic tumor vessels and several stromal cell types that, by interacting with each other and with tumor cells, affect TME remodeling, cancer growth and progression. Among the thyroid TME components, cancer-associated fibroblasts (CAFs) have gained more attention in the last years. Indeed, recent important evidence showed that thyroid CAFs strongly sustain thyroid cancer growth and progression by producing soluble factors and ECM proteins, which, in turn, deeply affect thyroid cancer cell behavior and aggressiveness. Hence, in this article, we describe the thyroid TME, focusing on the desmoplastic stromal reaction, which is a powerful indicator of thyroid cancer progression and an invasive growth pattern. In addition, we discuss the origins and features of the thyroid CAFs, their influence on thyroid cancer growth and progression, their role in remodeling the ECM and their immune-modulating functions. We finally debate therapeutic perspectives targeting CAFs.

Orthotopic and Heterotopic Murine Models of Pancreatic Cancer Exhibit Different Immunological Microenvironments and Different Responses to Immunotherapy

For decades, tumor-bearing murine models established using tumor cell lines have been the most commonly used models to study human cancers. Even though there are several studies reported that implant sites caused disparities in tumor behaviors, few of them illuminated the positional effect on immunotherapy. Herein, we describe surgical techniques for a novel orthotopic implantation of syngeneic pancreatic ductal adenocarcinoma (PDAC) tissue slices. This method has a high success modeling rate and stable growth kinetics, which makes it useful for testing novel therapeutics. Pathological examination indicated that the orthotopic tumor displayed poor vascularization, desmoplastic stromal reaction, and a highly immunosuppressive tumor microenvironment. This unique microenvironment resulted in limited response to PD1/CTLA4 blockade therapy and anti-MUC1 (αMUC1) CAR-T transfer treatment. To reverse the suppressive tumor microenvironment, we developed gene modified T-cells bearing a chimeric receptor in which activating receptor NKG2D fused to intracellular domains of 4-1BB and CD3ζ (NKG2D CAR). The NKG2D CAR-T cells target myeloid-derived suppressor cells (MDSCs), which overexpress Rae1 (NKG2D ligands) within the TME. Results indicated that NKG2D CAR-T cells eliminated MDSCs and improved antitumor activity of subsequently infused CAR-T cells. Moreover, we generated a bicistronic CAR-T, including αMUC1 CAR and NKG2D CAR separated by a P2A element. Treatment with the dual targeted bicistronic CAR-T cells also resulted in prolonged survival of orthotopic model mice. In summary, this study describes construction of a novel orthotopic PDAC model through implantation of tissue slices and discusses resistance to immunotherapy from the perspective of a PDAC microenvironment. Based on the obtained results, it is evident that elimination MDSCs by NKG2D CAR could rescue the impaired CAR-T cell activity.

CAF-Associated Paracrine Signaling Worsens Outcome and Potentially Contributes to Chemoresistance in Epithelial Ovarian Cancer.

BackgroundHigh-grade serous ovarian cancer (HGSOC) is the predominant and deadliest form of ovarian cancer. Some of its histological subtypes can be distinguished by frequent occurrence of cancer-associated myofibroblasts (CAFs) and desmoplastic stroma reaction (DSR). In this study, we want to explore the relationship between therapy outcome and the activity of CAF-associated signaling pathways in a homogeneous HGSOC patient collective. Furthermore, we want to validate these findings in a general Epithelial ovarian cancer (EOC) cohort.MethodsThe investigation cohort consists of 24 HGSOC patients. All of them were treated with platinum-based components and clinical follow-up was available. The validation cohort was comprised of 303 patients. Sequencing data (whole transcriptome) and clinical data were extracted from The Cancer Genome Atlas (TCGA). RNA of HGSOC patients was isolated using a Maxwell RSC instrument and the appropriate RNA isolation kit. For digital expression analysis a custom-designed gene panel was employed. All genes were linked to various DSR- and CAF- associated pathways. Expression analysis was performed on the NanoString nCounter platform. Finally, data were explored using the R programming environment (v. 4.0.3).ResultIn total, 15 CAF-associated genes were associated with patients’ survival. More specifically, 6 genes (MMP13, CGA, EPHA3, PSMD9, PITX2, PHLPP1) were linked to poor therapy outcome. Though a variety of different pathways appeared to be associated with therapy failure, many were related to CAF paracrine signaling, including MAPK, Ras and TGF-β pathways. Similar results were obtained from the validation cohort.DiscussionIn this study, we could successfully link CAF-associated pathways, as shown by increased Ras, MAPK and PI3K-Akt signaling to therapy failure (chemotherapy) in HGSOC and EOCs in general. As platinum-based chemotherapy has been the state-of-the-art therapy to treat HGSOC for decades, it is necessary to unveil the reasons behind resistance developments and poor outcome. In this work, CAF-associated signaling is shown to compromise therapy response. In the validation cohort, CAF-associated signaling is also associated with therapy failure in general EOC, possibly hinting towards a conserved mechanism. Therefore, it may be helpful to stratify HGSOC patients for CAF activity and consider alternative treatment options.

Abstract P3-15-03: Imaging and invasive lobular carcinoma: A survey study conducted by the lobular breast cancer alliance

Abstract Background: The Lobular Breast Cancer Alliance (LBCA) is an advocacy organization guided by an expert advisory board. Since it launched in 2017, LBCA has been committed to raising awareness of the distinct aspects of invasive lobular carcinoma (ILC) and promoting ILC research. While ILC is the second most common histological breast cancer type representing 15% of breast cancer diagnoses in the United States annually, it is understudied compared to the more common invasive ductal carcinoma (IDC). ILC tumor cell density and lack of desmoplastic stromal reaction can pose challenges to detection by physical examination and mammography. Many studies have shown decreased accuracy of breast imaging tools for ILC, and those with ILC are often diagnosed at later stages and require more extensive surgery. We sought to hear from members of the LBCA community with ILC about their experiences with detection of their tumors. Methods: LBCA conducted an anonymous survey online. A link to the survey on the SurveyMonkey platform was made available for two weeks in June. It was sent to LBCA newsletter subscribers and posted to the LBCA website and ILC social media groups. The survey questions included stage and size of tumors at initial diagnosis and after surgery, and questions related to imaging for advanced ILC. It specifically asked whether and what type of mammogram initially detected the ILC. Results: Within 24 hours over 1,000 people responded, and a total of 1,476 people previously diagnosed with ILC responded by survey close. The majority of participants, 1,052 (72%), were from the US, followed by participants from 35 additional countries. 170 (12%) reported having metastatic ILC. 1,035 (70%) of respondents were under 60 at diagnosis. Of respondents reporting tumor size and stage, 936 (67%) of respondents indicated having a tumor larger than 2cm at diagnosis, and 897 (63%) reported stage 2 or higher, including 73 (5%) reporting stage 4 at initial diagnosis. In the subset of participants initially diagnosed with stage 2 or higher, 410 (46%) of respondents reported the tumor was not detected by mammogram. In the subset of participants who noticed changes in their breasts prior to the diagnosis, 521 (79%) had tumors larger than 2cm, and 490 (74%) were diagnosed at stage 2 or higher. Of the 1,241 respondents who had no preoperative therapy, 531 (43%) had tumors larger than initially seen on imaging and/or 321 (26%) had more tumors than initially found on imaging. Discussion: To the best of our knowledge this represents the largest survey seeking to understand patients’ experience with imaging to diagnose or visualize ILC. The results from this community survey of patients diagnosed with invasive lobular carcinoma confirmed patients’ perceptions of mammography as an imperfect screening tool to detect ILC. Of note were the findings that mammography failed to visualize 46% of ILC cases at stage 2 or higher, and the majority of respondents had tumors diagnosed/detected at stage 2 or higher and at sizes greater than 2cm. Moreover, participants often reported that the actual tumor size on the final pathology specimen was larger than seen on diagnostic imaging, and a significant number of respondents were diagnosed de novo metastatic. In summary, our findings affirm patients’ perception about the limitations of diagnostic and monitoring imaging of ILC. Our impressively large response also demonstrates the urgent need from patients’ perspectives for more research on better methods for detecting and monitoring ILC to improve outcomes for patients diagnosed with lobular disease. Citation Format: Laurie B Hutcheson, Janice Axelrod, Colleen L Fitzwater, Maxine S Jochelson, Gitte H Joergensen, Theresa Langdon, Julia K Levine, Otto Metzger, Mason Mitchell-Daniels, Barbara F Neilsen. Imaging and invasive lobular carcinoma: A survey study conducted by the lobular breast cancer alliance [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P3-15-03.

Prognostic significance of tumour budding, tumour–stroma ratio and desmoplastic stromal reaction in gall bladder carcinoma

Aims and methodsThe prognostic role of tumour budding (TBd) and its interaction with the stromal microenvironment has gained a lot of attention recently, but remains unexplored in gall bladder cancer...