Tumor Fibroblasts Research Articles

Is the Thickness of the Margin Associated With Local Recurrence and Survival in Patients With Myxofibrosarcoma?

Myxofibrosarcoma (MFS) is a spectrum of aggressive soft tissue fibroblastic neoplasms characterized by variable myxoid stroma, pleomorphism, and a distinctive curved vascular pattern; these tumors are associated with a high likelihood of recurrence. Better local tumor control (a tumor-free margin) is believed to be important to minimize the risk of recurrence, but the effect of surgical resection margin status on local recurrence and survival in MFS is not as well-characterized as it might be. (1) Is margin width associated with local recurrence? (2) Is there a relationship between greater margin thickness and improved overall and disease-free survival (DFS)? (3) Is worsening French Federation of Cancer Centers grade associated with local recurrence and poorer overall survival? Using a database of patients with bone and soft tissue tumors at a tertiary university hospital, we retrospectively reviewed the medical records of 282 patients who had soft tissue sarcomas and who had been surgically treated by a multidisciplinary bone and soft tissue tumor care team between January 2010 and December 2021. Of these 282 patients, 38 were identified as having MFS. Patients who received surgical care for MFS outside our institution (unplanned resection) (four patients) and whose surgical margins were not reported as microscopic numerical data (10) were excluded from the analysis. We estimated survival and local recurrence and examined factors potentially influencing these outcomes. Patient demographics, tumor characteristics, surgical margin distance (in mm), and disease-related outcomes were recorded. The minimum follow-up was 3 months (median 41.5 months, range 3 to 128 months). Overall 1-year local recurrence-free rates were 66.7% (95% CI 50% to 88%). Patients with positive margins were more likely to have local recurrence than patients with negative margins (HR 10.91 [95% CI 2.61 to 45.66]; p = 0.001). Patients with an inadequate margin (positive margin or a negative margin of 1 mm or less) had a greater risk of local recurrence (HR 9.96 [95% CI 1.22 to 81.44]; p = 0.032). Patients with positive margins or margins less than or equal to 1 mm had worse 2-year local recurrence-free survival than did those with margins of greater than 1 mm (46.9% [95% CI 16% to 76%] versus 91.7% [95% CI 75% to 100%]; p = 0.005). The mean overall survival was 98 months (95% CI 77.2 to 118.8). The Kaplan-Meier overall 1-, 2- and 5-year estimated rates of survival were 88% (95% CI 75% to 100%), 79.2% (95% CI 64.5% to 97.2%), and 73.5% (95% CI 57.2 % to 94.5%), respectively. Positive surgical margins were associated with decreased overall survival (HR 6.96 [95% CI 1.39 to 34.89]; p = 0.018). There was a mean DFS time of 4.25 months (95% CI 0.92 to 7.59) in microscopically positive patients, 75.5 months (95% CI 37.47 to 113.53) in patients with margins 1 mm or less, and 118 months (95% CI 99.23 to 136.77) in patients with margins over 1 mm. There was a statistical difference between DFS times according to surgical margin classification (p < 0.001). With the numbers we had, we could not detect any difference between the histologic grades determined by the French Federation of Cancer Centers grading system in terms of local recurrence (HR 3.80 [95% CI 0.76 to 18.94]; p = 0.103) and overall survival (HR 6.91 [95% CI 0.79 to 60.13]; p = 0.080). Tumor size was the prognostic factor associated with a higher local recurrence rate among all factors analyzed as univariate (HR 1.18 [95% CI 1.05 to 1.32]; p = 0.004). A surgical procedure with a sufficient negative surgical margin distance appears to be associated with a lower proportion of patients who experience a local recurrence and is associated with overall patient survival. It is difficult to define what a sufficient margin is, but in our patients, it appears to be greater than 1 mm.Level of Evidence Level III, therapeutic study.

Establishment of Pancreatic Cancer-Derived Tumor Organoids and Fibroblasts From Fresh Tissue.

Tumor organoids are three-dimensional (3D) ex vivo tumor models that recapitulate the biological key features of the original primary tumor tissues. Patient-derived tumor organoids have been used in translational cancer research and can be applied to assess treatment sensitivity and resistance, cell-cell interactions, and tumor cell interactions with the tumor microenvironment. Tumor organoids are complex culture systems that require advanced cell culture techniques and culture media with specific growth factor cocktails and a biological basement membrane that mimics the extracellular environment. The ability to establish primary tumor cultures highly depends on the tissue of origin, the cellularity, and the clinical features of the tumor, such as the tumor grade. Furthermore, tissue sample collection, material quality and quantity, as well as correct biobanking and storage are crucial elements of this procedure. The technical capabilities of the laboratory are also crucial factors to consider. Here, we report a validated SOP/protocol that is technically and economically feasible for the culture of ex vivo tumor organoids from fresh tissue samples of pancreatic adenocarcinoma origin, either from fresh primary resected patient donor tissue or patient-derived xenografts (PDX). The technique described herein can be performed in laboratories with basic tissue culture and mouse facilities and is tailored for wide application in the translational oncology field.

5-ALA photodynamic ablation of fibroblastic soft-tissue tumors

Background: Fibroblastic soft-tissue tumors share enzymatic anomalies that result in excessive intracellular conversion of 5-aminolevulinic acid (5-ALA) to protoporphyrin IX, a photosensitizer which induces cellular apoptosis upon exposure to visible red light at a wavelength of 635 nm. We hypothesized that red light illumination of the surgical bed remaining after resection of fibroblastic tumors will result in destruction of microscopic tumor residua and may decrease the likelihood of local tumor recurrence. Methods: Twenty-four patients with desmoid tumors, solitary fibrous tumors (SFT), and dermatofibrosarcoma protuberans (DFSP) received oral 5-ALA prior to resection of their tumors. Following tumor resection, the exposed surgical bed was illuminated with red light at a wave length of 635 nm at a dose of 150 J/cm2 for 33 min. Results: Treatment with 5-ALA was associated with minor side effects that included nausea and transient elevation of transaminases. Local tumor recurrence was detected in 1 of the 10 patients with desmoid tumors who had not undergone any previous surgery, none in the 6 patients who had SFT and 1 of the 5 patients who had DFSP. Conclusions: 5-ALA photodynamic therapy of fibroblastic soft-tissue tumors may result in decreased likelihood of local tumor recurrence. It is associated with minimal side effects and should be considered as adjuvant to tumor resection in these cases.

The Role of Irisin/FNDC5 Expression and Its Serum Level in Breast Cancer.

Irisin (Ir) is an adipomyokine formed from fibronectin type III domain-containing protein 5 (FNDC5), which can be found in various cancer tissues. Additionally, FNDC5/Ir is suspected of inhibiting the epithelial-mesenchymal transition (EMT) process. This relationship has been poorly studied for breast cancer (BC). The ultrastructural cellular localizations of FNDC5/Ir were examined in BC tissues and BC cell lines. Furthermore, we compared serum levels of Ir with FNDC5/Ir expression in BC tissues. The aim of this study was to examine the levels of EMT markers, such as E-cadherin, N-cadherin, SNAIL, SLUG, and TWIST, and to compare their expression levels with FNDC5/Ir in BC tissues. Tissue microarrays with 541 BC samples were used to perform immunohistochemical reactions. Serum levels of Ir were assessed in 77 BC patients. We investigated FNDC5/Ir expression and ultrastructural localization in MCF-7, MDA-MB-231, and MDA-MB-468 BC cell lines and in the normal breast cell line (Me16c), which was used as the control. FNDC5/Ir was present in BC cell cytoplasm and tumor fibroblasts. FNDC5/Ir expression levels in BC cell lines were higher compared to those in the normal breast cell line. Serum Ir levels did not correlate with FNDC5/Ir expression in BC tissues but were associated with lymph node metastasis (N) and histological grade (G). We found that FNDC5/Ir correlated moderately with E-cadherin and SNAIL. Higher Ir serum level is associated with lymph node metastasis and increased grade of malignancy. FNDC5/Ir expression is associated with E-cadherin expression level.

Wide local excision, Mohs micrographic surgery, and reconstructive options for treatment of dermatofibrosarcoma protuberans of the breast: A retrospective case series from Mayo Clinic

BackgroundDermatofibrosarcoma protuberans (DFSP) of the breast is a dermal fibroblastic neoplasm requiring wide excisional margins due to recurrence rates ranging from 26 to 60%. The current literature on reconstructive options and utility of Mohs micrographic surgery for DFSP of the breast is scarce. We describe surgical management of DFSP of the breast at our institution with the largest case series reported to date.MethodsA retrospective review was performed of women who underwent surgery for DFSP of the breast at our institution between 1990 and 2019. Continuous data was summarized using mean, median, and range; categorical data was summarized with frequency count and percentage. Preoperative lesion size and postoperative defect size were evaluated using 2-sided Fisher exact test, and p-values < 0.05 were considered statistically significant.ResultsNine patients underwent wide local excision (WLE) with reconstruction including pedicled latissimus dorsi flaps (n = 2), local flap advancement (n = 2), mastectomy with implant (n = 1), oncoplastic breast reduction (n = 1), and skin grafts (n = 3). Nine underwent Mohs micrographic surgery (MMS) with complex primary closure. Mean postoperative maximum wound defect size for WLE was 10.8 cm versus 7.0 cm for MMS with no statistical significance (p = 0.77). Mean preoperative maximum lesion size for WLE was 6.4 cm versus 3.3 cm for MMS with no statistical significance (p = 0.07). Complications with WLE included wound dehiscence in three patients and seroma in one patient. No complications were reported with MMS and primary closure. Recurrence was reported in one WLE patient, which was successfully detected despite flap coverage and resected without complications. Median follow-up for the patients without recurrence was 5.0 years, with two patients in MMS cohort lost to follow-up. Five-year overall survival was 100%.ConclusionsMMS and WLE are both viable surgical options for managing DFSP of the breast. MMS could potentially minimize reconstructive needs due to smaller average defect size and result in fewer complications but may also result in asymmetry. Immediate flap reconstruction, especially in larger defects, can achieve excellent aesthetic outcomes for patients with DFSP of the breast without compromising detection of disease recurrence.

Advanced Cell Culture Models Illuminate the Interplay between Mammary Tumor Cells and Activated Fibroblasts.

The interaction between tumor cells and activated fibroblasts determines malignant features of desmoplastic carcinomas such as rapid growth, progression towards a metastatic phenotype, and resistance to chemotherapy. On one hand, tumor cells can activate normal fibroblasts and even reprogram them into CAFs through complex mechanisms that also involve soluble factors. Among them, transforming growth factor beta (TGF-β) and Platelet-Derived Growth Factor (PDGF) have an established role in the acquisition of pro-tumorigenic phenotypes by fibroblasts. On the other hand, activated fibroblasts release Interleukin-6 (IL-6), which increases tumor-cell invasiveness and chemoresistance. However, the interplay between breast cancer cells and fibroblasts, as well as the modes of action of TGF-β, PDGF, and IL-6, are difficult to investigate in vivo. Here, we validated the usage of advanced cell culture models as tools to study the interplay between mammary tumor cells and fibroblasts, taking mouse and human triple-negative tumor cells and fibroblasts as a case study. We employed two different settings, one permitting only paracrine signaling, the other both paracrine and cell-contact-based signaling. These co-culture systems allowed us to unmask how TGF-β, PDGF and IL-6 mediate the interplay between mammary tumor cells and fibroblasts. We found that the fibroblasts underwent activation induced by the TGF-β and the PDGF produced by the tumor cells, which increased their proliferation and IL-6 secretion. The IL-6 secreted by activated fibroblasts enhanced tumor-cell proliferation and chemoresistance. These results show that these breast cancer avatars possess an unexpected high level of complexity, which resembles that observed in vivo. As such, advanced co-cultures provide a pathologically relevant tractable system to study the role of the TME in breast cancer progression with a reductionist approach.

Modulation of EZH2 Activity Induces an Antitumoral Effect and Cell Redifferentiation in Anaplastic Thyroid Cancer.

Anaplastic thyroid cancer (ATC) is a rare and lethal form of thyroid cancer that requires urgent investigation of new molecular targets involved in its aggressive biology. In this context, the overactivation of Polycomb Repressive Complex 2/EZH2, which induces chromatin compaction, is frequently observed in aggressive solid tumors, making the EZH2 methyltransferase a potential target for treatment. However, the deregulation of chromatin accessibility is yet not fully investigated in thyroid cancer. In this study, EZH2 expression was modulated by CRISPR/Cas9-mediated gene editing and pharmacologically inhibited with EZH2 inhibitor EPZ6438 alone or in combination with the MAPK inhibitor U0126. The results showed that CRISPR/Cas9-induced EZH2 gene editing reduced cell growth, migration and invasion in vitro and resulted in a 90% reduction in tumor growth when EZH2-edited cells were injected into an immunocompromised mouse model. Immunohistochemistry analysis of the tumors revealed reduced tumor cell proliferation and less recruitment of cancer-associated fibroblasts in the EZH2-edited tumors compared to the control tumors. Moreover, EZH2 inhibition induced thyroid-differentiation genes' expression and mesenchymal-to-epithelial transition (MET) in ATC cells. Thus, this study shows that targeting EZH2 could be a promising neoadjuvant treatment for ATC, as it promotes antitumoral effects in vitro and in vivo and induces cell differentiation.

Cell-by-cell quantification of the androgen receptor in benign and malignant prostate leads to a better understanding of changes linked to cancer initiation and progression.

The androgen receptor (AR) plays a crucial role in the development and homeostasis of the prostate and is a key therapeutic target in prostate cancer (PCa). The gold standard therapy for advanced PCa is androgen deprivation therapy (ADT), which targets androgen production and AR signaling. However, resistance to ADT develops via AR-dependent and AR-independent mechanisms. As reports on AR expression patterns in PCa have been conflicting, we performed cell-by-cell AR quantification by immunohistochemistry in the benign and malignant prostate to monitor changes with disease development, progression, and hormonal treatment. Prostates from radical prostatectomy (RP) cases, both hormone-naïve and hormone-treated, prostate tissues from patients on palliative ADT, and bone metastases were included. In the normal prostate, AR is expressed in >99% of luminal cells, 51% of basal cells, and 61% of fibroblasts. An increase in the percentage of AR negative (%AR-) cancer cells along with a gradual loss of fibroblastic AR were observed with increasing Gleason grade and hormonal treatment. This was accompanied by a parallel increase in staining intensity of AR positive (AR+) cells under ADT. Staining AR with N- and C-terminal antibodies yielded similar results. The combination of %AR- cancer cells, %AR- fibroblasts, and AR intensity score led to the definition of an AR index, which was predictive of biochemical recurrence in the RP cohort and further stratified patients of intermediate risk. Lastly, androgen receptor variant 7 (ARV7)+ cells and AR- cells expressing neuroendocrine and stem markers were interspersed among a majority of AR+ cells in ADT cases. Altogether, the comprehensive quantification of AR expression in the prostate reveals concomitant changes in tumor cell subtypes and fibroblasts, emphasizing the significance of AR- cells with disease progression and palliative ADT.

Fibroma of tendon sheath: a clinicopathological and genetic analysis of 134 cases

Objective: To investigate the clinicopathological features, immunophenotypes and molecular genetics of fibroma of tendon sheath (FTS). Methods: One hundred and thirty-four cases of FTS or tenosynovial fibroma diagnosed in the Department of Pathology, West China Hospital, Sichuan University, Chengdu, China from January 2008 to April 2019 were selected. The clinical and histologic features of these cases were retrospectively reviewed. Immunohistochemistry, fluorescence in situ hybridization (FISH) and reverse transcription-polymerase chain reaction (RT-PCR) were performed on the above cases. Results: There were a total of 134 cases of FTS, including 67 males and 67 females. The patients' median age was 38 years (ranged from 2 to 85 years). The median tumor size was 1.8 cm (ranged from 0.1 to 6.8 cm). The most common site was the upper extremity (76/134, 57%). Follow-up data was available in 28 cases and there was no detectable recurrence. Classic FTS (114 cases) were well-defined and hypocellular. A few spindle-shaped fibroblasts were scattered in the dense collagenous sclerotic stroma. Characteristically elongated slit-like spaces or thin-walled vessels were observed. Most of cellular FTSs (20 cases) were well-defined and the area with increased cellularity of the spindle cells coexisted with classic FTS. There were occasional mitotic figures, but no atypical mitotic figures. Immunohistochemistry was performed in 8 cases of classic FTS and most cases were positive for SMA (5/8). Immunohistochemistry was also performed in 13 cases of cellular FTS and showed 100% positive rate for SMA. FISH was conducted on 20 cases of cellular FTS and 32 cases of classical FTS. USP6 gene rearrangement was found in 11/20 of cellular FTS. Among 12 cases of CFTS with nodular fasciitis (NF)-like morphological feature, 7 cases showed USP6 gene rearrangement. The rearrangement proportion of USP6 gene in cellular FTS without NF-like morphological features was 4/8. By contrast, 3% (1/32) of the classic FTS showed USP6 gene rearrangement. RT-PCR was performed in those cases with detected USP6 gene rearrangement and sufficient tissue samples for RT-PCR. The MYH9-USP6 fusion gene was detected in 1 case (1/8) of the cellular FTSs, while no target fusion partner was detected in the classic FTS. Conclusions: FTS is a relatively rare benign fibroblastic or myofibroblastic tumor. Our study and recent literature find that some of the classic FTS also show USP6 gene rearrangements, suggesting that classical FTS and cellular FTS are likely to be at different stages of the same disease (spectrum). FISH for USP6 gene rearrangement may be used as an important auxiliary diagnostic tool in distinguishing FTS from other tumors.

Abstract 1364: Profiling tumor microenvironment for therapeutic intervention to soft-tissue sarcomas

Abstract Soft-tissue sarcomas (STS) are mesenchymal tumors having remarkably diverse histological features. Genomic studies reported that STS have low frequencies of genetic mutation, but often harboring copy number variations (CNVs). For STS, CNVs frequently happen in essential tumor suppressor genes and oncogenic transcriptional genes, neither of which are easily “druggable” targets. Moreover, in concordance with low tumor mutation burden rate, STS show low immunogenicity with less cytotoxic TIL in the TME and is unfortunately linked to a lower responsive rate to PD1/PD-L1 ICB. Using a syngeneic mouse UPS model, we previously found that tumor-associated macrophages (TAM) in the TME are promoting the growth of sarcoma. We further identified specific tumor microenvironmental elements contributing to the pro-tumorigenicity of TAMs, which represent a promising novel clinical target for myeloid-related intervention. Therefore, we hypothesize that exploration of TME is beneficial to identify clinically promising targets for STS. To study the TME in STS, we performed bulk RNA-seq and scRNA-seq on human sarcoma samples which covered different histological subtypes of STS and compare them to normal samples. Our results show that, unlike carcinomas originating from epithelial cells, extracellular matrix (ECM)-related genes are highly upregulated in both cancer-associated fibroblast (CAF) and sarcoma tumor cells. Further analysis show that upregulation of these ECM genes is associated with a worse survival outcome in sarcoma patients. In order to further explore the STS and TME orchestra, we established several syngeneic mouse models according to the genetic aberrations reported in TCGA data. We are currently characterizing these syngeneic mouse models to find out whether they recapitulate human STS samples. Our goal is to utilize these mouse models to study the relationship of ECM-related genes in both CAF and tumor cells, find out the effect of upregulated ECM-related genes on the immune compartment in TME and search for potential therapeutic interventions. Citation Format: Jin-Fen Xiao, Marina Broz, Roberta Piras, Kristin Ishaya, Emily Ko, Jlenia Guarnerio. Profiling tumor microenvironment for therapeutic intervention to soft-tissue sarcomas [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 1364.

Abstract 5424: A deep learning approach (AI) which accurately identifies breast tumor cells, tumor infiltrating lymphocytes (TILS) and fibroblasts from H&amp;E slides

Abstract Background: Histopathology assessments of cancers require highly skilled pathologists, are labor intensive and prone to errors without proper training or fatigue. Machine learning can assist pathologists by increasing efficiency and minimizing individual variability. This study adapted a deep learning model to reliably identify 3 cell types from triple negative breast cancers fixed on H&amp;E slides and verified performance with an expert pathologist. Methods: We apply the U-Net architecture to analyse pathology slides fixed with triple negative breast cancer (TNBC) tissue. The public dataset NuCLS was used for training. Semantic segmentation was used to identify single cells of 3 types: tumor cells, fibroblasts, and tumor infiltrating lymphocyte (TILs). For validation, the pathologist annotated 8 random H&amp;E tiles. The 3 cell types accuracy for NuCLS and the model was evaluated by our pathologist. Results: Overall, there was a 73% agreement with the pathologists (Pathologist vs. NuCLS). A set of 1,555 (90%) TNBC slides were used for training and 173 for validation (10%; unseen data). Table 1 outlines the accuracy metrics for each cell type and for each comparison. Compared to our pathologist, the model accurately identified TILs (62%), followed by fibroblasts (42%) and lastly tumor cells (26%). A significant source of discrepancy was variation in labeled single cell boundaries. The model was better at identifying TILs. The pathologist took 1.5 hours to annotate 8 tiles for the 3 cells and our model 644ms. Conclusion: It is possible to develop a deep learning model to identify breast cancer cells, fibroblasts and TILs from H&amp;E stained slides, with similar accuracy levels as a trained pathologist. The model performed better than a pathologist in identifying TILs, but both struggled with fibroblasts. Accuracy of 71% overall and 87% for TILs, motivates expansion to further datasets and other cancer types. Table 1. - Accuracy metrics. Quality assessment8 H&amp;E tiles Pathologist vs. NuCLS U-Net model vs. NuCLs U-Net model vs. Pathologist U-Net model vs. NuCLs (Full) Background 86% 71% 62% 73% Tumour 43% 35% 26% 71% Fibroblast 45% 39% 42% 31% TILs 34% 96% 62% 87% Overall 73% 74% 61% 71% Citation Format: Luiz Augusto Zillmann da Silva, Alistair R. Williams, Aidan Kubeyev, Andrea Giorni, Jordan Laurie, Prabu Sivasubramaniam, Matthew Foster, Matthew Griffiths, Uzma Asghar. A deep learning approach (AI) which accurately identifies breast tumor cells, tumor infiltrating lymphocytes (TILS) and fibroblasts from H&amp;E slides. [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 5424.

Mediastinal Myxofibrosarcoma Harboring Loss-of-Function MSH2 Variant in a Patient With Lynch Syndrome: A Case Report and Literature Review.

Myxofibrosarcoma is a malignant fibroblastic neoplasm that commonly arises in the extremities, with mediastinum being a very rare location. The development of sarcomas is uncommon in patients with Lynch syndrome. We present a Lynch syndrome patient with synchronous cecal adenocarcinoma and mediastinal myxofibrosarcoma with both harboring the same loss-of-function MSH2 alteration (c.2634 + 1G > A splice region variant). Metastatic myxofibrosarcoma in the left chest wall developed 6 months after the initial diagnosis. The clinical presentation, imaging findings, histopathology, and molecular studies along with differential diagnoses are presented and discussed.

Desmoid-type fibromatosis in splenic hilum with rupture of lienal artery aneurism in a 17-year-old male

Desmoid Fibromatosis (DF) is locally aggressive benign fibroblastic or myofibroblastic tumor with no metastatic potential, but has high recurrence rate. The symptoms vary according to tumor location, abdominal or extra-abdominal.

Myxofibrosarcoma of Head and Neck Region in a Patient with Follicular Carcinoma of Thyroid

Myxofibrosarcoma (MFS) is a malignant fibroblastic tumour with variable myxoid stroma, cellular pleomorphism and curvilinear vascular pattern. It is commonly seen in late adult life, peaks in the seventh decade and usually occurs in the lower extremities. MFS of head and of the neck is extremely rare and accounts for approximately 1% of all head and neck neoplasms. A fifty two year old lady with follicular carcinoma thyroid treated six years ago, presented with a rapidly increasing neck swelling in the left supraclavicular fossa. Fine needle aspiration cytology of the cervical node was suggestive of metastatic carcinoma and she underwent left posterolateral neck dissection. Histopathology and immunohistochemistry was suggestive of high grade myxofibrosarcoma with negative margins. She was treated with adjuvant radiotherapy and has been under regular follow up for 6 months.

ER+, HER2- advanced breast cancer treated with taselisib and fulvestrant: genomic landscape and associated clinical outcomes.

Taselisib is a potent β-sparing phosphatidylinositol 3-kinase (PI3K) inhibitor that, with endocrine therapy, improves outcomes in phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA)-mutated (PIK3CAmut) advanced breast cancer. To understand alterations associated with response to PI3K inhibition, we analysed circulating tumour DNA (ctDNA) from participants enrolled in the SANDPIPER trial. Participants were designated as either PIK3CAmut or PIK3CA no mutation was detected (NMD) per baseline ctDNA. The top mutated genes and tumour fraction estimates identified were analysed for their association with outcomes. In participants with PIK3CAmut ctDNA treated with taselisib + fulvestrant, tumour protein p53 (TP53; encoding p53) and fibroblast growth factor receptor 1 (FGFR1) alterations were associated with shorter progression-free survival (PFS) compared to participants with NMD in these genes. Conversely, participants with PIK3CAmut ctDNA harbouring a neurofibromin 1 (NF1)alteration or high baseline tumour fraction estimate experienced improved PFS upon treatment with taselisib + fulvestrant compared to placebo + fulvestrant. Broadly, alterations in oestrogen receptor (ER), PI3K and p53 pathway genes were associated with resistance to taselisib + fulvestrant in participants with PIK3CAmut ctDNA. Altogether, we demonstrated the impact of genomic (co-)alterations on outcomes with one of the largest clinico-genomic datasets of ER+, HER2-, PIK3CAmut breast cancer patients treated with a PI3K inhibitor.

Clinic and anatomic aspects of verification and monitoring of various types of equine sarcoid in the western regions of Ukraine

Sarcoid is the most common skin tumor reported worldwide in equids, donkeys, zebras, and mules of all ages and sexes. The bovine papillomavirus (BPV) plays an important role in the pathogenesis of the disease. Globally, the incidence of equine sarcoid ranges from 12 % to 67 % of all skin neoplasms. Unfortunately, no attention is paid to the study of sarcoid epizootology in Ukraine. There are no reports on the clinical types of neoplasms, their pathohistological characteristics, and treatment methods. Our study aimed to describe the clinical manifestations of different anatomical types of equine sarcoid, ranging from small single lesions to multiple aggressive fibroblastic tumors, which were registered in horses from private farms and stables in the western regions of Ukraine during 2019–2022. According to its macroscopic features, sarcoid was divided into six clinical and anatomical types: hidden, verrucous, nodular, fibroblastic, mixed, and malignant. In general, 1012 horses were examined, of which 328 animals were clinically diagnosed with sarcoid. Neoplasms were localized in different parts of the body and belonged to different sarcoid types.The smallest number of neoplasms was detected in the area of the head around the eyes in the form of single nodules (2.3 %), on the neck (5.4 %), and limbs (14.8 %). Most of the multiple focal sarcoid tumors were localized in the chest area (26.5 %) and in the area of the abdominal wall and groin (51 %). Both depigmentation and hyperkeratinization of the affected areas were the typical morphological features of all sarcoid types. Hidden sarcoid was characterized by lesions of the skin’s superficial layers with well-defined areas of alopecia. Verrucous sarcoid was mainly localized in the neck and groin areas with the formation of irregularly shaped and large skin thickenings characterized by damage to the deep dermis layers. The third type of sarcoid is nodular, which was visually manifested in the form of mobile spherical subcutaneous nodules of small sizes. They were usually localized in the groin, prepuce, inner thighs, and eyelids. Fibroblastic type is one of the most aggressive equine sarcoids. It is characterized by rapid growth with the formation of massive tumors on the leg with an ulcerated surface and marked vascularization. Mixed sarcoid combined verrucous, fibroblastic and nodular types with predominant signs a particular type of tumor. Malignant form of sarcoid was relatively rare but particularly aggressive. Tumors spread along the course of lymphatic vessels with the formation of nodes and ulcers. This sarcoid was detected only in two horses. In general, it should be noted that sarcoid is not a fatal neoplasm for horses. However, its location, size, and the possibility of progression to a more aggressive form commonly leads to deterioration of the exterior and culling of thoroughbred animals.

Fibromatosis of the breast; another diagnosis of cancer like lesions: About 4 cases reports

Introduction and importanceDesmoid fibromatosis is a benign monoclonal fibroblastic tumor that was first reported in 1832 and arises from musculoaponeurotic structures. It is an uncommon tumor that can mimic breast carcinoma in both hits clinical and radiological presentation.This entity mainly affects patients in their fourth and fifth decades of life. Case presentationOur four patients were of young women in their 2nd and 3rd decade that consulted for masses of the breast.These lesions were suspicious for malignancy in both their clinical and radiological features despite not having any family history of breast carcinoma. Clinical discussionOnly histology provides the diagnosis, it is locally invasive, non-metastatic but with a high potential of recurrence and the sole treatment is complete and wide local excision with clear margins. ConclusionAlthough it is a rare clinical scenario, the presence of an unusual breast or chest wall tumor such as a desmoid tumor should be considered when obvious breast changes and history do not correlate with routine diagnostic measures.

Clinicopathologic and genetic characterization of angiofibroma of soft tissue: a study of 12 cases including two cases with AHRR::NCOA3 gene fusion.

Angiofibroma of soft tissue (AFST) is a benign tumour characterised by prominent arborizing blood vessels throughout the lesion. Approximately two-thirds of AFST cases were reported to have AHRR::NCOA2 fusion, and only two cases have been reported to have other gene fusions: GTF2I::NCOA2 or GAB1::ABL1. Although AFST is included in fibroblastic and myofibroblastic tumours in the World Health Organization's 2020 classification, histiocytic markers, especially CD163, have been reported to be positive in almost all examined cases, and it still remains the possibility of a fibrohistiocytic nature of the tumour. Therefore, we aimed to clarify the genetic and pathological spectrum of AFST and identify whether histiocytic marker-positive cells were true neoplastic cells. We evaluated 12 AFST cases, which included 10 cases with AHRR::NCOA2 and two with AHRR::NCOA3 fusions. Pathologically, nuclear palisading, which has not been reported in AFST, was detected in two cases. Furthermore, one tumour resected by additional wide resection revealed severe infiltrative growth. Immunohistochemical analysis indicated varying levels of desmin-positive cells in nine cases, whereas CD163- and CD68-positive cells were diffusely distributed in all 12 cases. We also performed double immunofluorescence staining and immunofluorescence in situ hybridisation in four resected cases with >10% desmin-positive tumour cells. The results suggested that the CD163-positive cells differed from desmin-positive cells with AHRR::NCOA2 fusion in all four cases. Our findings suggested that AHRR::NCOA3 could be the second most frequent fusion gene, and histiocytic marker-positive cells are not genuine neoplastic cells in AFST.

Evaluation of tumour necrosis factor alpha-stimulated gene-6 and fibroblast growth factor-2 levels in patients diagnosed with multi-system inflammatory syndrome in children.

Investigations are still ongoing about the pathophysiology of multi-system inflammatory syndrome in children, which can progress with serious morbidity and mortality after COVID-19 infection. In this study, we aimed to investigate whether fibroblast growth factor-2 and tumour necrosis factor alpha-stimulated gene-6 levels play a role in the diagnosis of the disease and on cardiac involvement. Twenty-three patients (11 girls, 12 boys) and 26 healthy controls (10 girls, 16 boys) were included in the study. The mean age of the patient and control group was 8.45 ± 2.43 and 10.73 ± 4.27 years, respectively. There was no difference between the fibroblast growth factor-2 and tumour necrosis factor alpha-stimulated gene-6 levels of the patient and control groups. When the patients with myocardial involvement in the patient group were compared with the patients without myocardial involvement in terms of fibroblast growth factor-2 and tumour necrosis factor alpha-stimulated gene-6 levels, no difference was found between these groups. The correlation of fibroblast growth factor-2 and tumour necrosis factor alpha-stimulated gene-6 levels with other laboratory parameters was investigated in the patient group. Fibroblast growth factor-2 was moderately inversely correlated with white blood cell count (r = -0.541, p = 0.008), absolute neutrophil count (r = -0.502, p = 0.015) and C-reactive protein (r = -0.528, p = 0.010). Fibroblast growth factor-2 was strongly inversely correlated with erythrocyte sedimentation rate (r = -0.694, p =<0.001). Our data show that fibroblast growth factor-2 and tumour necrosis factor alpha stimulated gene-6 do not provide sufficient information about diagnosis and cardiac involvement in multi-system inflammatory syndrome in children.

Microfluidic Droplet-Assisted Fabrication of Vessel-Supported Tumors for Preclinical Drug Discovery.

High-fidelity in vitro tumor models are important for preclinical drug discovery processes. Currently, the most commonly used model for in vitro drug testing remains the two-dimensional (2D) cell monolayer. However, the natural in vivo tumor microenvironment (TME) consists of extracellular matrix (ECM), supporting stromal cells and vasculature. They not only participate in the progression of tumors but also hinder drug delivery and effectiveness on tumor cells. Here, we report an integrated engineering system to generate vessel-supported tumors for preclinical drug screening. First, gelatin-methacryloyl (GelMA) hydrogel was selected to mimic tumor extracellular matrix (ECM). HCT-116 tumor cells were encapsulated into individual micro-GelMA beads with microfluidic droplet technique to mimic tumor-ECM interactions in vitro. Then, normal human lung fibroblasts were mingled with tumor cells to imitate the tumor-stromal interaction. The tumor cells and fibroblasts reconstituted in the individual GelMA microbead and formed a biomimetic heterotypic tumor model with a core-shell structure. Next, the cell-laden beads were consociated into a functional on-chip vessel network platform to restore the tumor-tumor microenvironment (TME) interaction. Afterward, the anticancer drug paclitaxel was tested on the individual and vessel-supported tumor models. It was demonstrated that the blood vessel-associated TME conferred significant additional drug resistance in the drug screening experiment. The reported system is expected to enable the large-scale fabrication of vessel-supported heterotypic tumor models of various cellular compositions. It is believed to be promising for the large-scale fabrication of biomimetic in vitro tumor models and may be valuable for improving the efficiency of preclinical drug discovery processes.