Suitability of ultrasound-guided fine-needle aspiration biopsy for transcriptome sequencing of the canine prostate

Abstract

Ultrasound-guided fine-needle aspiration (US-FNA) biopsy is a widely used minimally invasive sampling procedure for cytological diagnosis. This study investigates the feasibility of using US-FNA samples for both cytological diagnosis and whole transcriptome RNA-sequencing analysis (RNA-Seq), with the ultimate aim of improving canine prostate cancer management. The feasibility of the US-FNA procedure was evaluated intra vitam on 43 dogs. Additionally, aspirates from 31 euthanised dogs were collected for standardising the procedure. Each aspirate was separated into two subsamples: for cytology and RNA extraction. Additional prostate tissue samples served as control for RNA quantity and quality evaluation, and differential expression analysis. The US-FNA sampling procedure was feasible in 95% of dogs. RNA isolation of US-FNA samples was successfully performed using phenol-chloroform extraction. The extracted RNA of 56% of a subset of US-FNA samples met the quality requirements for RNA-Seq. Expression analysis revealed that only 153 genes were exclusively differentially expressed between non-malignant US-FNAs and tissues. Moreover, only 36 differentially expressed genes were associated with the US-FNA sampling technique and unrelated to the diagnosis. Furthermore, the gene expression profiles clearly distinguished between non-malignant and malignant samples. This proves US-FNA to be useful for molecular profiling.

Transcriptome of Two Canine Prostate Cancer Cells Treated With Toceranib Phosphate Reveals Distinct Antitumor Profiles Associated With the PDGFR Pathway

“…Besides, we validated our results comparing our DEGs found in treated PC cell lines with gene expression profile of primary tumors (23). Interestingly, we found gene expression in opposite direction to that found in tumor tissues and these could be interesting genes to understand the modulation of toceranib phosphate therapy in these canine prostate carcinomas.…”

“…We found 1,412 up and 668 downregulated genes shared by the primary tumors from biopsy and needle-aspiration (23). By comparing the gene expression profile of primary tumors with the treated cell lines we observed that PC1 has 59 overlapping genes while PC2 has 11 overlapping genes ( Figure 8A).…”

“…To evaluate the expression profile of PC1 and PC2 in primary tumors we downloaded the RNAseq data from GSE122916 study available at GEO (Gene Expression Omnibus) database (23). We then performed differential expression analysis using the NetworkAnalyst 3.0 software (24)(25)(26).…”

Contrast-Enhanced Ultrasound Imaging of Prostate Gland in Neutered Dogs

“…Histological evaluation by ultrasound-guided fine needle aspiration (FNA) has been shown to be the most accurate test for detecting PN. Nevertheless, due to the low contrast and specificity existing between malignant areas and normal tissues on ultrasound images, nontargeted 12-site systematic biopsies covering the whole prostate gland are often performed to detect neoplasia, causing unnecessary pain to the patient [18][19][20]. Therefore, due to the lack of specific tumor ultrasonographic patterns, it is of great importance to develop innovative imaging techniques to increase the sensitivity and specificity of prostate cancer screening at an early stage, which may become very helpful to collect fewer and targeted samples, thereby reducing pain and stress.…”

Metformin and sodium dichloroacetate effects on proliferation, apoptosis, and metabolic activity tested alone and in combination in a canine prostate and a bladder cancer cell line

“…Canine prostate adenocarcinoma and transitional cell carcinoma are characterized by a local and invasive growth pattern and are clinically difficult to differentiate in dogs. Both tumors are associated with a poor prognosis due to late diagnosis, metastatic and invasive behavior, and limited therapeutic options [ 1 – 3 ]. Moreover, canine prostate adenocarcinoma is refractory to androgen deprivation therapy, which is the first-line therapy for prostate cancer in humans [ 4 , 5 ].…”