Automated Ki-67 Quantification of Immunohistochemical Staining Image of Human Nasopharyngeal Carcinoma Xenografts

To generate a reliable preclinical model system exhibiting the molecular features of salivary adenoid cystic carcinoma (ACC) whose biology is still unclear due to the paucity of stable cell cultures. To develop new in vitro and in vivo models of ACC, the techniques of organoid culture and patient-derived tumor xenograft (PDX), which have attracted attention in other malignancies in recent years, were applied. Tumor specimens from surgically resected salivary ACC were proceeded for the preparation of PDX and organoid culture. The orthotopic transplantation of patient-derived or PDX-derived organoids was demonstrated into submandibular glands of NSG mice and those histology was evaluated. PDX-derived organoid cells were evaluated for the presence of MYB-mediated fusion genes and proceeded for in vitro drug sensitivity assay. Human ACC-derived organoids were successfully generated in three-dimensional culture and confirmed the ability of these cells to form tumors by orthotopic injection. Short-term organoid cell cultures from two individual ACC PDX tumors were also established that maintain the characteristic MYBL1 translocation and histological features of the original parent and PDX tumors. Finally, the establishment of drug sensitivity tests on these short-term cultured cells was confirmed using three different agents. This is the first to report an approach for the generation of human ACC-derived organoids as in vitro and in vivo cancer models, providing insights into understanding of the ACC biology and creating personalized therapy design for patients with ACC. K E Y W O R D S drug sensitivity test, organoid culture, orthotopic transplantation, patient-derived xenograft (PDX) model, salivary adenoid cystic carcinoma 1 | INTRODUCTION Adenoid cystic carcinomas (ACC) is one of the most common types of salivary gland malignancies, representing approximately 1% of all head and neck malignancies and about 10% to 15% of all salivary gland cancers. 1,2 Salivary ACC generally represents as slow growth tumor, however, also

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“…19 Briefly, after anesthetizing, an incision of about Ki-67 index was then calculated as described previously. 20…”

Section: Transplantation Of Organoids and Pdx-derived Organoidsmentioning

confidence: 99%

Establishment of PDX‐derived salivary adenoid cystic carcinoma cell lines using organoid culture method

Takada

Aizawa

Sano

et al. 2020

Intl Journal of Cancer

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“…However, the statistical significance and reliability of the proposed approach on limited number of tissue sample is to be justified. The adequate sample size N could be calculated using paired t ‐test (Shi et al ., ),

N = {([], ((), Z_{α / 2} + Z_{β}) σ / δ)}^{2},

where, δ is the allowable error limit, which can be set at 5%, σ is the population standard deviation of the difference value in the proposed approach from expert score,

Z_{α / 2}

is the two‐tailed critical value of normal standard distribution and

Z_{β}

is considered regardless of one or two tailed test with α and β are set to 0.05 and 0.1, respectively.…”

Section: Workflowmentioning

confidence: 97%

Section: Sample Size Calculationmentioning

confidence: 99%

AutoIHC‐scoring: a machine learning framework for automated Allred scoring of molecular expression in ER‐ and PR‐stained breast cancer tissue

Tewary

Arun

Ahmed

et al. 2017

Journal of Microscopy

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In prognostic evaluation of breast cancer Immunohistochemical (IHC) markers namely, oestrogen receptor (ER) and progesterone receptor (PR) are widely used. The expert pathologist investigates qualitatively the stained tissue slide under microscope to provide the Allred score; which is clinically used for therapeutic decision making. Such qualitative judgment is time-consuming, tedious and more often suffers from interobserver variability. As a result, it leads to imprecise IHC score for ER and PR. To overcome this, there is an urgent need of developing a reliable and efficient IHC quantifier for high throughput decision making. In view of this, our study aims at developing an automated IHC profiler for quantitative assessment of ER and PR molecular expression from stained tissue images. We propose here to use CMYK colour space for positively and negatively stained cell extraction for proportion score. Also colour features are used for quantitative assessment of intensity scoring among the positively stained cells. Five different machine learning models namely artificial neural network, Naïve Bayes, K-nearest neighbours, decision tree and random forest are considered for learning the colour features using average red, green and blue pixel values of positively stained cell patches. Fifty cases of ER- and PR-stained tissues have been evaluated for validation with the expert pathologist's score. All five models perform adequately where random forest shows the best correlation with the expert's score (Pearson's correlation coefficient = 0.9192). In the proposed approach the average variation of diaminobenzidine (DAB) to nuclear area from the expert's score is found to be 7.58%, as compared to 27.83% for state-of-the-art ImmunoRatio software.

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“…The overall survival rate of patients with NPC has been increasing; however the prognosis of patients with advanced NPC remains poor and the 5-year survival rate of such patients is only 30–40% ( 6 , 7 ). Tumor recurrence and distant metastasis are the primary factors affecting the prognosis of patients with NPC ( 8 ).…”

Section: Introductionmentioning

confidence: 99%

Claudin1 promotes the proliferation, invasion and migration of nasopharyngeal carcinoma cells by upregulating the expression and nuclear entry of β‑catenin

Xiao

Zhao

et al. 2018

Exp Ther Med

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The aim of the present study was to measure the expression of Claudin (CLDN) 1 in nasopharyngeal carcinoma (NPC) and to determine its biological function and mechanism of action. Reverse transcription-quantitative polymerase chain reaction and western blotting were performed to measure the expression of CLDN1 mRNA and protein, respectively, in the immortalized human nasopharyngeal epithelial cell line NP69 and NPC-TW01 cells. Subsequently, small interfering RNA against CLDN1 and the LV-GFP-PURO-CLDN1 lentivirus were transfected into NPC-TW01 cells. Western blotting was used to determine the effects of CLDN1 down- and upregulation on the expression of the epithelial mesenchymal transition (EMT) markers E-cadherin and vimentin. In addition, the effect of CLDN1 on the expression of β-Catenin was determined. The results demonstrated that levels of CLDN1 mRNA and protein in NPC cells were significantly higher than in NP69 cells. Furthermore, the downregulation of CLDN1 inhibited the proliferation, invasion and migration of NPC-TW01 cells. The results of western blotting demonstrated that the downregulation of CLDN1 resulted in the upregulation of E-cadherin and inhibition of vimentin in NPC-TW01 cells. By contrast, the overexpression of CLDN1 resulted in the downregulation of E-cadherin and upregulation of vimentin in NPC-TW01 cells. The downregulation of β-catenin attenuated the cancer-promoting effect of CLDN1 on NPC-TW01 cells, whereas the upregulation of β-catenin reversed the tumor-suppressing effect of CLDN1 downregulation on NPC-TW01 cells. The results of the present study therefore demonstrate that CLDN1 expression is elevated in NPC cells. As an oncogene, CLDN1 promotes the proliferation, invasion and migration of NPC cells by upregulating the expression and nuclear entry of β-catenin.

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Automated Ki-67 Quantification of Immunohistochemical Staining Image of Human Nasopharyngeal Carcinoma Xenografts

Cited by 43 publications

References 22 publications

Establishment of PDX‐derived salivary adenoid cystic carcinoma cell lines using organoid culture method

Establishment of PDX‐derived salivary adenoid cystic carcinoma cell lines using organoid culture method

AutoIHC‐scoring: a machine learning framework for automated Allred scoring of molecular expression in ER‐ and PR‐stained breast cancer tissue

Claudin1 promotes the proliferation, invasion and migration of nasopharyngeal carcinoma cells by upregulating the expression and nuclear entry of β‑catenin

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