Quantitative analysis of the BRAF V595E mutation in plasma cell-free DNA from dogs with urothelial carcinoma

Tagawa, Michihito; Tambo, Naomi; Maezawa, Masaki; Tomihari, Mizuki; Watanabe, Kenichi; Inokuma, Hisashi; Miyahara, Koshiro

doi:10.1371/journal.pone.0232365

Cited by 22 publications

(27 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Second, cfDNA is a very sensitive diagnostic biomarker for inflammation; therefore, it is difficult to completely rule out the possibility of elevated cfDNA levels in non-specific inflammation in vivo . Real-time PCR techniques with high specificity can be used to determine the underlying inflammatory mechanisms ( 24 – 27 ). Third, the observation period was short (up to 60 weeks), and each individual case had a different time frame.…”

Section: Discussionmentioning

confidence: 99%

Cell-Free DNA as a Diagnostic and Prognostic Biomarker in Dogs With Tumors

et al. 2021

View full text Add to dashboard Cite

Cell-free DNA (cfDNA) is derived from apoptosis/necrosis, active cellular secretion, and lysis of circulating cancer cells or micrometastases. In humans, cfDNA is widely used in cancer diagnosis, but veterinary research has yet to be actively conducted to establish it as a cancer biomarker. This retrospective study analyzed cfDNA levels in samples collected from dogs with neoplastic disease (n = 38), clinically ill dogs without neoplasia (n = 47), and healthy dogs (n = 35). cfDNA levels and clinical data were compared among groups, and prognostic analyses were performed within the neoplastic group. Furthermore, continual cfDNA measurements were performed during the chemotherapy of six dogs with lymphoma. Dogs with neoplasia showed significantly higher cfDNA concentrations than dogs without neoplasm, and the cfDNA oncentration in the lymphoid neoplasia group was significantly elevated among all neoplastic groups. Dogs with neoplasia and a plasma cfDNA concentration above 1,247.5 μg/L had shorter survival rates than those with levels below this threshold (26.5 vs. 86.1%, respectively, P < 0.05). In cases with complete remission in response to chemotherapy, the cfDNA concentration was significantly decreased compared with the first visit, whereas the cfDNA concentration was increased in cases with disease progression or death. Interestingly, a significant correlation was found between lymph node diameter and cfDNA concentration in dogs with multicentric lymphoma (R2 = 0.26, P < 0.01). These data suggest that changes in cfDNA concentration could be used as a diagnostic biomarker for canine neoplasia. Furthermore, increased plasma DNA levels might be associated with shorter survival time, and cfDNA concentrations may reflect the response to chemotherapy.

show abstract

Section: Discussionmentioning

confidence: 99%

Cell-Free DNA as a Diagnostic and Prognostic Biomarker in Dogs With Tumors

et al. 2021

View full text Add to dashboard Cite

show abstract

“…In a recent study of canine mammary carcinoma, ctDNA detection was demonstrated using digital PCR assays specific to somatic genomic rearrangements in 4 dogs (Beck et al 2013). In another study, ctDNA detection was demonstrated in 8 of 11 dogs with urothelial carcinoma using a real-time PCR assay for a recurrent somatic mutation in BRAF (Tagawa et al 2020). Earlier, we demonstrated detection of ctDNA in 2 of 6 dogs with pulmonary adenocarcinoma using a digital PCR assay for a recurrent somatic mutation in ERBB2 (Lorch et al 2019).…”

Section: Introductionmentioning

confidence: 87%

“…However, published experience with ctDNA analysis in dogs is currently sparse. Nonetheless, limited and small-scale published studies have suggested total cell-free DNA concentrations are higher in dogs with malignant tumors compared to those with benign disease or healthy controls (Schaefer et al 2007;Beffagna et al 2017;Tagawa et al 2020). In a recent study of canine mammary carcinoma, ctDNA detection was demonstrated using digital PCR assays specific to somatic genomic rearrangements in 4 dogs (Beck et al 2013).…”

Section: Introductionmentioning

confidence: 99%

Feasibility and promise of circulating tumor DNA analysis in dogs with naturally-occurring sarcoma

Favaro

Stewart²,

McDonald

et al. 2020

Preprint

View full text Add to dashboard Cite

Comparative studies of naturally-occurring canine cancers have provided new insight into many areas of cancer research. The inclusion of pet dogs in the development and validation of circulating tumor DNA (ctDNA) diagnostics may be uniquely informative for human translation for many reasons, including: high incidence of certain spontaneous cancers, repeated access to blood and tumor from the same individuals during the course of disease progression, and molecular heterogeneity of naturally-occurring cancers in dogs. Here, we present a feasibility study of ctDNA analysis performed in 9 healthy dogs and 39 dogs with either benign or malignant splenic tumors (hemangiosarcoma) using shallow whole genome sequencing (sWGS) of cell-free DNA. To enable detection and quantification of ctDNA using sWGS, we adapted two informatic approaches and compared their performance for the canine genome. At presentation, mean ctDNA tumor fraction in dogs with malignant splenic tumors was 11.2%, significantly higher than dogs with benign lesions (3.2%; p 0.001), achieving an AUC of 0.84. ctDNA tumor fraction was 14.3% and 9.0% in dogs with metastatic and localized disease, respectively although this difference was not statistically significant (p 0.227). In paired analysis, ctDNA fraction decreased from 11.0% to 7.9% after resection of malignant tumors (p 0.047). Our results demonstrate that ctDNA analysis is feasible in dogs with hemangiosarcoma using a cost-effective approach such as sWGS. Future studies are underway to validate these findings, and further evaluate the role of ctDNA to assess burden of disease and treatment response during drug development.

show abstract

“…One example already in diagnostic use is the previously mentioned BRAF mutation for the detection of UC. A recent advance of this approach has been the demonstration that mutated BRAF can be detected in blood samples from these patients (Tagawa et al 2020). A prognostic example is PARR in combination with quantitative PCR for monitoring residual disease burden in canine lymphoma patients.…”

Section: Identification Of Novel Biomarkers and Therapeutic Targets To Predict Ideal Therapy Choicesmentioning

confidence: 99%

Molecular pathology in the cancer clinic – where are we now and where are we headed?

Guillén

Smallwood

Killick

2021

J of Small Animal Practice

View full text Add to dashboard Cite

Molecular pathology is a developing sub-microscopic discipline of pathology that studies the effects of molecular variations and mutations on disease processes. The ultimate goal of molecular pathology in cancer is to predict risk, facilitate diagnosis and improve prognostication based on a complete understanding of the biological impact of specific molecular variations, mutations and dysregulations. This knowledge will provide the basis for customised cancer treatment, so-called precision medicine. Rapid developments in genomics have placed this field at the forefront of clinical molecular pathology and there are already a number of well-established genetic tests available for clinical use including PCR of antigen receptor rearrangement and KIT mutational analysis. Moving beyond tests assessing a single gene, there are significant research efforts utilising genomics to predict cancer risk, forecast aggressive behaviour and identify druggable mutations and therapeutic biomarkers. Researchers are also investigating the use of circulating cells and nucleic acid for clinically useful low morbidity genomic assessments. If we are to realise the full potential of molecular pathology and precision medicine there are a number of challenges to overcome. These include developing our understanding of the underlying biology (in particular intra-tumoural heterogeneity), methodological standardisation of assays, provision of adequate infrastructure and production of novel therapeutics backed by high-quality clinical data supporting the precision medicine approach. The era of molecular pathology holds the potential to revolutionise veterinary cancer care, but its impact on clinical practice will depend upon the extent to which the inherent challenges can be overcome.

show abstract

Quantitative analysis of the BRAF V595E mutation in plasma cell-free DNA from dogs with urothelial carcinoma

Cited by 22 publications

References 25 publications

Cell-Free DNA as a Diagnostic and Prognostic Biomarker in Dogs With Tumors

Cell-Free DNA as a Diagnostic and Prognostic Biomarker in Dogs With Tumors

Feasibility and promise of circulating tumor DNA analysis in dogs with naturally-occurring sarcoma

Molecular pathology in the cancer clinic – where are we now and where are we headed?

Contact Info

Product

Resources

About