Digital image analysis and machine learning-assisted prediction of neoadjuvant chemotherapy response in triple-negative breast cancer.

BACKGROUND: Pathological complete response (pCR) is associated with favorable prognosis in patients with triple-negative breast cancer (TNBC). However, only 30-40% of TNBC patients treated with neoadjuvant chemotherapy (NAC) show pCR, while the remaining 60-70% show residual disease (RD). The role of the tumor microenvironment in NAC response in patients with TNBC remains unclear. In this study, we developed a machine learning-based two-step pipeline to distinguish between various histological components in hematoxylin and eosin (H&E)-stained whole slide images (WSIs) of TNBC tissue biopsies and to identify histological features that can predict NAC response. METHODS: H&E-stained WSIs of treatment-naïve biopsies from 85 patients (51 with pCR and 34 with RD) of the model development cohort and 79 patients (41 with pCR and 38 with RD) of the validation cohort were separated through a stratified eightfold cross-validation strategy for the first step and leave-one-out cross-validation strategy for the second step. A tile-level histology label prediction pipeline and four machine-learning classifiers were used to analyze 468,043 tiles of WSIs. The best-trained classifier used 55 texture features from each tile to produce a probability profile during testing. The predicted histology classes were used to generate a histology classification map of the spatial distributions of different tissue regions. A patient-level NAC response prediction pipeline was trained with features derived from paired histology classification maps. The top graph-based features capturing the relevant spatial information across the different histological classes were provided to the radial basis function kernel support vector machine (rbfSVM) classifier for NAC treatment response prediction. RESULTS: The tile-level prediction pipeline achieved 86.72% accuracy for histology class classification, while the patient-level pipeline achieved 83.53% NAC response (pCR vs. RD) prediction accuracy of the model development cohort. The model was validated with an independent cohort with tile histology validation accuracy of 83.59% and NAC prediction accuracy of 81.01%. The histological class pairs with the strongest NAC response predictive ability were tumor and tumor tumor-infiltrating lymphocytes for pCR and microvessel density and polyploid giant cancer cells for RD. CONCLUSION: Our machine learning pipeline can robustly identify clinically relevant histological classes that predict NAC response in TNBC patients and may help guide patient selection for NAC treatment.

Digital image analysis and machine learning-assisted prediction of neoadjuvant chemotherapy response in triple-negative breast cancer.

Background: Pathological complete response (pCR) is associated with favorable prognosis in patients with triple-negative breast cancer (TNBC). However, only 30-40% of TNBC patients treated with neoadjuvant chemotherapy (NAC) show pCR, while the remaining 60-70% show residual disease (RD). The role of the tumor microenvironment (TME) in NAC response in patients with TNBC remains unclear. In this study, we developed a machine learning-based two-step pipeline to distinguish between various histological components in hematoxylin and eosin (H&E)-stained whole slide images (WSIs) of TNBC tissue biopsies and to identify histological features that can predict NAC response. Methods: H&E-stained WSIs of treatment-naïve biopsies from 85 patients (51 with pCR and 34 with RD) were separated through a stratified 8-fold cross validation strategy for the first step and leave one out cross validation strategy for the second step. A tile-level histology label prediction pipeline and four machine learning classifiers were used to analyze 468,043 tiles of WSIs. The best-trained classifier used 55 texture features from each tile to produce a probability profile during testing. The predicted histology classes were used to generate a histology classification map of the spatial distributions of different tissue regions. A patient-level NAC response prediction pipeline was trained with features derived from paired histology classification maps. The top graph-based features capturing the relevant spatial information across the different histological classes were provided to the radial basis function kernel support vector machine (rbfSVM) classifier for NAC treatment response prediction. Results: The tile-level prediction pipeline achieved 86.72% accuracy for histology class classification, while the patient-level pipeline achieved 83.53% NAC response (pCR vs. RD) prediction accuracy. The histological class pairs with the strongest NAC response predictive ability were tumor and tumor tumor-infiltrating lymphocytes for pCR and microvessel density and polyploid giant cancer cells for RD. Conclusion: Our machine learning pipeline can robustly identify clinically relevant histological classes that predict NAC response in TNBC patients and may help guide patient selection for NAC treatment.

Stromal Expression of CD10 in Breast Carcinoma and Its Association with Known Prognostic Factors-A Tissue Microarray-Based Study.

Background Breast cancer is an epithelial malignancy; however, stroma plays a key role with its stimulatory and inhibitory factors in modulating tumor invasion and metastasis. CD10, a matrix metalloproteinase, is known to regulate cell adhesion, migration and helps in determining the progression of tumor. This knowledge helps to identify specific signals that promote growth, dedifferentiation, invasion, metastasis and serve as target for better therapeutic management. Objectives The aim of this study was to estimate frequency of expression of stromal CD10 and assess its prognostic significance in breast carcinomas by correlating with known prognostic factors. Materials and Methods Morphological parameters of 62 cases of carcinoma breast were studied on H&E (hematoxylin and eosin) stained sections and expressions of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2/neu), and CD10 on manually constructed tissue microarray sections by immunohistochemistry (IHC). Staining pattern, percentage of stained cells, and intensity of stains were evaluated and IHC scoring of all markers was done. CD10 scores were correlated with the known prognostic factors (ER, PR, and HER2/neu). A p -value less than 0.05 was considered as significant. Results Stromal expression of CD10 was found in 82.3% of cases and it was significantly associated with increasing tumor size ( p = 0.012), increasing tumor grade ( p = 0.001), lymph node metastasis ( p = 0.018), necrosis ( p = 0.008), lymphovascular invasion ( p = 0.008), ER negativity ( p = 0.001), PR negativity( p = 0.007), HER 2 positivity ( p = 0.012), triple-negative molecular subtypes ( p = 0.001), and poor prognostic groups ( p = 0.01). Conclusion CD10 can be used as an independent prognostic stromal marker and this will help to envisage new therapeutic strategies.

PLK1 and AURKB phosphorylate survivin differentially to affect proliferation in racially distinct triple-negative breast cancer.

Protein diversity due to alternative mRNA splicing or post-translational modifications (PTMs) plays a vital role in various cellular functions. The mitotic kinases polo-like kinase 1 (PLK1) and Aurora B (AURKB) phosphorylate survivin, an inhibitor of apoptosis (IAP) family member, thereby regulating cell proliferation. PLK1, AURKB, and survivin are overexpressed in triple-negative breast cancer (TNBC), an aggressive breast cancer subtype. TNBC is associated with high proliferative capacity, high rates of distant metastasis, and treatment resistance. The proliferation-promoting protein survivin and its activating kinases, PLK1 and AURKB, are overexpressed in TNBC. In this study, we investigated the role of survivin phosphorylation in racial disparities in TNBC cell proliferation. Analysis of TCGA TNBC data revealed higher expression levels of PLK1 (P = 0.026) and AURKB (P = 0.045) in African Americans (AAs; n = 41) than in European Americans (EAs; n = 86). In contrast, no significant racial differences in survivin mRNA or protein levels were observed. AA TNBC cells exhibited higher p-survivin levels than EA TNBC cells. Survivin silencing using small interfering RNAs significantly attenuated cell proliferation and cell cycle progression in AA TNBC cells, but not in EA TNBC cells. In addition, PLK1 and AURKB inhibition with volasertib and barasertib significantly inhibited the growth of AA TNBC xenografts, but not of EA TNBC tumors. These data suggest that inhibition of PLK1 and AURKB suppresses cell proliferation and tumor growth, specifically in AA TNBC. These findings suggest that targeting survivin phosphorylation may be a viable therapeutic option for AA patients with TNBC.

Polyploid giant cancer cell characterization: New frontiers in predicting response to chemotherapy in breast cancer.

Although polyploid cells were first described nearly two centuries ago, their ability to proliferate has only recently been demonstrated. It also becomes increasingly evident that a subset of tumor cells, polyploid giant cancer cells (PGCCs), play a critical role in the pathophysiology of breast cancer (BC), among other cancer types. In BC, PGCCs can arise in response to therapy-induced stress. Their progeny possess cancer stem cell (CSC) properties and can repopulate the tumor. By modulating the tumor microenvironment (TME), PGCCs promote BC progression, chemoresistance, metastasis, and relapse and ultimately impact the survival of BC patients. Given their pro- tumorigenic roles, PGCCs have been proposed to possess the ability to predict treatment response and patient prognosis in BC. Traditionally, DNA cytometry has been used to detect PGCCs.. The field will further derive benefit from the development of approaches to accurately detect PGCCs and their progeny using robust PGCC biomarkers. In this review, we present the current state of knowledge about the clinical relevance of PGCCs in BC. We also propose to use an artificial intelligence-assisted image analysis pipeline to identify PGCC and map their interactions with other TME components, thereby facilitating the clinical implementation of PGCCs as biomarkers to predict treatment response and survival outcomes in BC patients. Finally, we summarize efforts to therapeutically target PGCCs to prevent chemoresistance and improve clinical outcomes in patients with BC.

Targeted Inhibition of Anti-Inflammatory Regulator Nrf2 Results in Breast Cancer Retardation In Vitro and In Vivo.

Nuclear factor erythroid-2 related factor-2 (Nrf2) is an oxidative stress-response transcriptional activator that promotes carcinogenesis through metabolic reprogramming, tumor promoting inflammation, and therapeutic resistance. However, the extension of Nrf2 expression and its involvement in regulation of breast cancer (BC) responses to chemotherapy remain largely unclear. This study determined the expression of Nrf2 in BC tissues (n = 46) and cell lines (MDA-MB-453, MCF-7, MDA-MB-231, MDA-MB-468) with diverse phenotypes. Immunohistochemical (IHC)analysis indicated lower Nrf2 expression in normal breast tissues, compared to BC samples, although the difference was not found to be significant. However, pharmacological inhibition and siRNA-induced downregulation of Nrf2 were marked by decreased activity of NADPH quinone oxidoreductase 1 (NQO1), a direct target of Nrf2. Silenced or inhibited Nrf2 signaling resulted in reduced BC proliferation and migration, cell cycle arrest, activation of apoptosis, and sensitization of BC cells to cisplatin in vitro. Ehrlich Ascites Carcinoma (EAC) cells demonstrated elevated levels of Nrf2 and were further tested in experimental mouse models in vivo. Intraperitoneal administration of pharmacological Nrf2 inhibitor brusatol slowed tumor cell growth. Brusatol increased lymphocyte trafficking towards engrafted tumor tissue in vivo, suggesting activation of anti-cancer effects in tumor microenvironment. Further large-scale BC testing is needed to confirm Nrf2 marker and therapeutic capacities for chemo sensitization in drug resistant and advanced tumors.

Repositioning of Itraconazole for the Management of Ocular Neovascularization Through Surface-Modified Nanostructured Lipid Carriers.

Retinopathy is one of the most common complications of diabetes. Approximately 80% of patients with diabetes history for over 10 years suffer from some degree of diabetic retinopathy (DR). Currently available treatments include use of antivascular endothelial growth factor-165 (VEGF165) agents or steroids. However, they are very expensive, involve an invasive procedure that is painful, and show ocular and systemic complications. Currently, the focus for treatment of such disorders has shifted from new drug discovery to repositioning of available drugs because of the cost and time consumption involved in the former. Working on this strategy, itraconazole (ITR) was selected for treatment of DR due to its potent unutilized antiangiogenic activity for the management of DR. An attempt was made to develop a topical, noninvasive nanostructured lipid carrier (NLC) owing to the potential to carry entrapped drug across the membranes. ITR-NLCs were prepared using high-pressure homogenization by applying Box-Behnken design for optimization. Surface of NLCs was modified by chitosan (CS) coating. ITR-NLCs were examined for antiangiogenic potential and their VEGF165 targeting efficiency. Drug-loaded NLC showed desired particle size, zeta potential, and polydispersity index. In VEGF-induced DR rats, ITR and CS-ITR-NLCs were found to exhibit an antineovascularization effect by targeting VEGF165. The developed CS-ITR-NLC proved to be an effective topical therapy for management of DR, offering the advantages of cost-effectiveness, higher patient compliance, and better tolerance.

Efficacy of Transcutaneous Electrical Nerve Stimulation in the Treatment of Overactive Bladder.

INTRODUCTION: Overactive Bladder (OAB) accounts for 40-70% cases of incontinence. The etiology is unknown though detrusor instability is found in urodynamic evaluation of almost all cases. Detrusor instability or hyperreflexia can be inhibited by direct inhibition of impulses in the pre-ganglionic afferent neuron or by inhibition of bladder pre-ganglionic neurons of the efferent limb of micturition reflex. Transcutaneous Electrical Nerve Stimulation (TENS) is based on the gate control theory of abolishing the local micturition reflex arc. AIM: To assess the effectiveness and safety of TENS in idiopathic OAB. MATERIALS AND METHODS: It is a prospective experimental study to evaluate the effectiveness of TENS v/s placebo in reducing OAB symptoms. (n1=20, n2 =20). Ten treatment sessions (5 sessions/week) of 30 minutes, were conducted. RESULTS: There was a significant improvement in Overactive Bladder Symptom Scores (OABSS) in TENS group and 2 patients were completely dry following TENS therapy. CONCLUSION: In elderly women, patients with OAB where other co-medications have their own anticholinergic side effects and impairment of cognition is a concern, TENS can be a useful intervention. TENS units are safe, economical and easily commercially available.

Cytological diagnosis of superficial acral fibromyxoma: A case report.

Superficial acral fibromyxoma (SAF) is a rare, distinctive benign soft tissue lesion that often involves the fingers and toes, with the great toe being the most frequently affected site. We report a case of SAF diagnosed by fine needle aspiration cytology and confirmed by histopathology. The pre-operative cytological diagnosis will help the surgeon to plan for a wider excision that prevents recurrence.

Study of computed tomography-guided fine needle aspiration cytology of thoracic lesions.

BACKGROUND: Thoracic lesions include a variety of benign and malignant lesions of lung, pleura, chest wall and mediastinum. Transthoracic fine needle aspiration cytology (TFNAC) is a well established technique for work up of thoracic lesions. Computed tomography (CT) has extended the use of FNAC, because it is accurate for localization, needle puncture and above all it permits evaluation of lesions less than 1 cm. This diagnostic modality has a high sensitivity, specificity and is of relatively low cost. AIMS: To assess the role of CT-guided FNAC in the diagnosis of thoracic lesions. MATERIALS AND METHODS: Eighty three patients with various thoracic lesions were evaluated by CT guided FNAC. The cytologic findings were compared with cell blocks whenever available. RESULTS: Conclusive opinion was offered on cytology smears in 80 patients. Lesions of the lung were the most common. Neoplastic lesions in our study accounted for 65% of cases. The sensitivity and specificity of the study were 93.33% and 100%, respectively. CONCLUSION: CT-guided TFNAC is a low cost, safe, minimally invasive and accurate diagnostic procedure with high sensitivity and specificity and when interpreted in conjunction with clinical and radiological data can prevent some of the pitfalls in diagnosis.

Juvenile hyaline fibromatosis.

Juvenile hyaline fibromatosis is a rare, autosomal-recessive disease characterized by papular and nodular skin lesions, gingival hyperplasia, joint contractures and bone involvement in variable degrees. It is a connective tissue disorder with aberrant synthesis of glycosaminoglycans by fibroblasts. We report a 5-year-old female born of first-degree consanguineous marriage who presented with multiple, recurrent, painless, variable-sized nodules. Fine needle aspiration cytology smears and the subsequent histopathological examination from the nodules showed benign spindle cells in a Periodic acid Schiff-positive myxoid background. The disease has a relentlessly progressive course, with most patients surviving only up to the 4(th) decade. As of now, there is no specific treatment for this disorder. Genetic counseling is essential to explain to parents about a 25% chance of having a diseased baby in any pregnancy. With the gene being mapped recently, techniques for antenatal diagnosis are likely to be established.

The cytology of micropapillary variant of colloid carcinoma of breast: A report of two cases.

Colloid carcinoma (pure mucinous carcinoma) is an uncommon variant of breast carcinoma with distinctive cytological and histological features. These tumors are characterised by islands of tumor cells floating in a sea of abundant extracellular mucin. We present the cytology of two cases of colloid carcinoma occurring in 80-year-old and 45-year-old females. The fine-needle aspiration cytology helps to subcategorize the tumor type, thereby enhancing the knowledge about the distinctive cytological features of special and uncommon variants of breast carcinoma, their course and prognosis. A distinctive micropapillary variant of pure mucinous carcinoma which is rarely described, is represented in one of the cases. Also we report, here, colloid carcinoma in a female of reproductive age, a relatively uncommon occurrence.

The cytology of molluscum contagiosum mimicking skin adnexal tumor.

Molluscum contagiosum is a cutaneous viral infection presenting as multiple, umbilicated papules and vesicles. The cytology of molluscum contagiosum in an 11-year-old boy, which presented atypically as a solitary nodule over the right cheek, mimicking a skin adnexal tumor is reported here. Fine needle aspiration cytology plays a vital role in establishing the correct diagnosis of clinically unsuspected cases, and hence, the proper management of such lesions. The cytology of molluscum contagiosum is characterized by the presence of numerous large intracytoplasmic basophilic bodies that push the host nucleus to the periphery, giving a signet ring appearance to the superficial epidermal cells.

Osseous differentiation in cystosarcoma phyllodes - diagnosed by fine needle aspiration cytology.

Osseous differentiation within a phyllodes tumor is extremely rare. Cytological and histological findings of a case of malignant phyllodes tumor with osseous differentiation are presented. A 45-year-old female had a malignant phyllodes tumor with osseous stroma diagnosed by fine needle aspiration cytology. The cytological findings were representative of the histological features. The diagnosis of these tumors preoperatively is important in planning the most appropriate treatment. It is also important to follow up these patients postoperatively for long periods for recurrence and metastasis.

Aggressive digital papillary adenocarcinoma diagnosed by fine needle aspiration cytology.

Aggressive digital papillary adenocarcinoma is a rare variant of eccrine sweat gland malignancy with a propensity for metastases and recurrence. We report a 45-year-old female with aggressive digital papillary adenocarcinoma diagnosed by fine needle aspiration cytology (FNAC). The cytological findings were representative of the histological features. The recognition of aggressive digital papillary adenocarcinoma as a distinct clinicopathological eccrine sweat gland neoplasm is important because of the potential for aggressive local growth and distant metastasis. FNAC plays an important role in the preoperative diagnosis and management of these lesions.

The cytology of giant solitary trichoepithelioma.

Giant solitary trichoepithelioma (GST) is a rare trichogenic tumor, which may present as a pigmented lesion. An 80-year-old man was diagnosed to have giant solitary trichoepithelioma on fine-needle aspiration cytology. The cytological findings represented the histological features. The recognition of GST is important because of its close resemblance to basal cell carcinoma and other skin adnexal tumors - clinically, cytologically and histologically.

Primary mucinous carcinoma of the skin: a rare tumor in the gluteal region.

Primary mucinous carcinoma (PMC) of the skin is a rare adnexal tumor of sweat gland origin. A case report is presented of a 50-year-old female who presented with a gluteal mass, which was diagnosed as an injection abscess. Following incision and drainage, the incision site persisted as a non-healing ulcer. An edge biopsy of the lesion revealed mucinous carcinoma of the skin. Investigations excluded the possibility of a metastatic mucinous carcinoma. Thus, the lesion in the gluteal region was diagnosed as PMC of the skin, a rare site of occurrence.