Detection of PIK3CA hotspot mutations in canine mammary tumors using droplet digital PCR: tissue validation and liquid biopsy feasibility.
Dogs
Animals
Class I Phosphatidylinositol 3-Kinases
/ genetics
Female
Liquid Biopsy
/ methods
Mammary Neoplasms, Animal
/ genetics
Mutation
Polymerase Chain Reaction
/ methods
Dog Diseases
/ genetics
High-Throughput Nucleotide Sequencing
/ methods
Feasibility Studies
Sensitivity and Specificity
Biomarkers, Tumor
/ genetics
PIK3CA mutations
Canine mammary tumors
Circulating tumor DNA
Droplet digital PCR
Liquid biopsy
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
26 Oct 2024
26 Oct 2024
Historique:
received:
26
07
2024
accepted:
16
10
2024
medline:
27
10
2024
pubmed:
27
10
2024
entrez:
27
10
2024
Statut:
epublish
Résumé
Domestic dogs (Canis lupus familiaris) serve as valuable translational models for human cancer research due to their biological similarities. Canine mammary tumors (CMTs), frequently diagnosed in female dogs, share various characteristics with human breast cancers. This study investigates the PIK3CA (H1047R) mutation in CMTs using droplet digital PCR (ddPCR) and explores the potential of liquid biopsy for non-invasive detection. We analyzed 80 formalin-fixed, paraffin-embedded (FFPE) CMT tissue samples and compared ddPCR results with next-generation sequencing (NGS) data, achieving high concordance. Plasma and serum samples were also assessed for mutation concordance with tissue results. Our findings indicate a higher frequency of the PIK3CA (H1047R) mutations in benign and grade I malignant CMTs compared to more aggressive malignancies. The ddPCR assay demonstrated high sensitivity and specificity, with plasma testing showing 78.6% sensitivity and 87.5% specificity, and serum testing showing 66.7% sensitivity and 90.0% specificity. These results highlight the viability of liquid biopsy as a minimally invasive method for monitoring PIK3CA mutations in canine patients. The study suggests that liquid biopsy techniques hold significant promise for improving the early detection and monitoring of canine cancers, warranting further research to refine these methods and explore their applications in canine cancer diagnostics and treatment.
Identifiants
pubmed: 39462049
doi: 10.1038/s41598-024-76820-0
pii: 10.1038/s41598-024-76820-0
doi:
Substances chimiques
Class I Phosphatidylinositol 3-Kinases
EC 2.7.1.137
Biomarkers, Tumor
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
25587Subventions
Organisme : National Research Foundation of Korea
ID : 2016M3A9B6903437
Informations de copyright
© 2024. The Author(s).
Références
Rowell, J. L., McCarthy, D. O. & Alvarez, C. E. Dog models of naturally occurring cancer. Trends Mol. Med. 17, 380–388 (2011).
doi: 10.1016/j.molmed.2011.02.004
pubmed: 21439907
pmcid: 3130881
Pang, L. Y. & Argyle, D. J. Using naturally occurring tumours in dogs and cats to study telomerase and cancer stem cell biology. Biochim. Biophys. Acta. 1792, 380–391 (2009).
doi: 10.1016/j.bbadis.2009.02.010
pubmed: 19254761
Heer, E. et al. Global burden and trends in premenopausal and postmenopausal breast cancer: A population-based study. Lancet Glob. Health 8, e1027–e1037 (2020).
doi: 10.1016/S2214-109X(20)30215-1
pubmed: 32710860
Salas, Y., Márquez, A., Diaz, D. & Romero, L. Epidemiological study of mammary tumors in female dogs diagnosed during the period 2002–2012: A growing animal health problem. PLoS One 10, e0127381 (2015).
doi: 10.1371/journal.pone.0127381
pubmed: 25992997
pmcid: 4436381
Gray, M. et al. Naturally-occurring canine mammary tumors as a translational model for human breast cancer. Front. Oncol. 10, 617 (2020).
doi: 10.3389/fonc.2020.00617
pubmed: 32411603
pmcid: 7198768
Diaz Jr, L. A. & Bardelli, A. Liquid biopsies: Genotyping circulating tumor DNA. J. Clin. Oncol. 32, 579–586 (2014).
doi: 10.1200/JCO.2012.45.2011
pubmed: 24449238
Flory, A. et al. Clinical validation of a next-generation sequencing-based multi-cancer early detection liquid biopsy blood test in over 1,000 dogs using an independent testing set: The CANcer detection in dogs (CANDiD) study. PLoS One 17, e0266623 (2022).
doi: 10.1371/journal.pone.0266623
pubmed: 35471999
pmcid: 9041869
Olmedillas-López, S., Olivera-Salazar, R., García-Arranz, M. & García-Olmo, D. Current and emerging applications of droplet digital PCR in oncology: An updated review. Mol. Diagn. Ther. 26, 61–87 (2022).
doi: 10.1007/s40291-021-00562-2
pubmed: 34773243
Zhao, J. J. et al. The p110alpha isoform of PI3K is essential for proper growth factor signaling and oncogenic transformation. Proc. Natl. Acad. Sci. U. S. A. 103, 16296–16300 (2006).
doi: 10.1073/pnas.0607899103
pubmed: 17060635
pmcid: 1637576
Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature 490, 61–70 (2012).
doi: 10.1038/nature11412
Nik-Zainal, S. et al. Landscape of somatic mutations in 560 breast cancer whole-genome sequences. Nature 534, 47–54 (2016).
doi: 10.1038/nature17676
pubmed: 27135926
pmcid: 4910866
Meyer, D. S. et al. Luminal expression of PIK3CA mutant H1047R in the mammary gland induces heterogeneous tumors. Cancer Res. 71, 4344–4351 (2011).
doi: 10.1158/0008-5472.CAN-10-3827
pubmed: 21482677
Bader, A. G., Kang, S., Zhao, L. & Vogt, P. K. Oncogenic PI3K deregulates transcription and translation. Nat. Rev. Cancer. 5, 921–929 (2005).
doi: 10.1038/nrc1753
pubmed: 16341083
Arafeh, R. & Samuels, Y. PIK3CA in cancer: The past 30 years. Semin. Cancer Biol. 59, 36–49 (2019).
doi: 10.1016/j.semcancer.2019.02.002
pubmed: 30742905
Liu, P., Cheng, H., Roberts, T. M. & Zhao, J. J. Targeting the phosphoinositide 3-kinase pathway in cancer. Nat. Rev. Drug Discov. 8, 627–644 (2009).
doi: 10.1038/nrd2926
pubmed: 19644473
pmcid: 3142564
Martini, M., De Santis, M. C., Braccini, L., Gulluni, F. & Hirsch, E. PI3K/AKT signaling pathway and cancer: An updated review. Ann. Med. 46, 372–383 (2014).
doi: 10.3109/07853890.2014.912836
pubmed: 24897931
Lee, K. H., Hwang, H. J., Noh, H. J., Shin, T. J. & Cho, J. Y. Somatic mutation of PIK3CA (H1047R) is a common driver mutation hotspot in canine mammary tumors as well as human breast cancers. Cancers (Basel) 11, 2006 (2019).
doi: 10.3390/cancers11122006
pubmed: 31842489
Kim, T. M. et al. Cross-species oncogenic signatures of breast cancer in canine mammary tumors. Nat. Commun. 11, 3616 (2020).
doi: 10.1038/s41467-020-17458-0
pubmed: 32680987
pmcid: 7367841
Alsaihati, B. A. et al. Canine tumor mutational burden is correlated with TP53 mutation across tumor types and breeds. Nat. Commun. 12, 4670 (2021).
doi: 10.1038/s41467-021-24836-9
pubmed: 34344882
pmcid: 8333103
Kim, S. H. et al. Dysregulation of PI3K/Akt/PTEN pathway in canine mammary tumor. Animals (Basel) 11, 2079 (2021).
doi: 10.3390/ani11072079
pubmed: 34359206
McEvoy, A. C. et al. Droplet digital PCR for mutation detection in formalin-fixed, paraffin-embedded melanoma tissues: A comparison with sanger sequencing and pyrosequencing. J. Mol. Diagn. 20, 240–252 (2018).
doi: 10.1016/j.jmoldx.2017.11.009
pubmed: 29305225
Zhang, B. et al. Comparison of droplet digital PCR and conventional quantitative PCR for measuring EGFR gene mutation. Exp. Ther. Med. 9, 1383–1388 (2015).
doi: 10.3892/etm.2015.2221
pubmed: 25780439
pmcid: 4353752
Arendt, M. L. et al. PIK3CA is recurrently mutated in canine mammary tumors, similarly to in human mammary neoplasia. Sci. Rep. 13, 632 (2023).
doi: 10.1038/s41598-023-27664-7
pubmed: 36635367
pmcid: 9837039
Martínez-Sáez, O. et al. Frequency and spectrum of PIK3CA somatic mutations in breast cancer. Breast Cancer Res. 22, 45 (2020).
doi: 10.1186/s13058-020-01284-9
pubmed: 32404150
pmcid: 7222307
Van Keymeulen, A. et al. Reactivation of multipotency by oncogenic PIK3CA induces breast tumour heterogeneity. Nature. 525, 119–123 (2015).
doi: 10.1038/nature14665
pubmed: 26266985
Bader, A. G., Kang, S. & Vogt, P. K. Cancer-specific mutations in PIK3CA are oncogenic in vivo. Proc. Natl. Acad. Sci. U. S. A. 103, 1475–1479 (2006).
doi: 10.1073/pnas.0510857103
pubmed: 16432179
pmcid: 1360603
Saad, E. S. A. et al. A comparative review of mixed mammary tumors in mammals. J. Mammary Gland Biol. Neoplasia. 24, 125–137 (2019).
doi: 10.1007/s10911-018-9422-2
pubmed: 30488318
Didelot, A. et al. Multiplex picoliter-droplet digital PCR for quantitative assessment of DNA integrity in clinical samples. Clin. Chem. 59, 815–823 (2013).
doi: 10.1373/clinchem.2012.193409
pubmed: 23403697
Rhrissorrakrai, K., Utro, F., Levovitz, C. & Parida, L. Lesion shedding model: Unraveling site-specific contributions to ctDNA. Brief. Bioinform. 24, bbad059 (2023).
doi: 10.1093/bib/bbad059
pubmed: 36869848
pmcid: 10025438
Sorenmo, K. U. et al. Canine mammary gland tumours; a histological continuum from benign to malignant; clinical and histopathological evidence. Vet. Comp. Oncol. 7, 162–172 (2009).
doi: 10.1111/j.1476-5829.2009.00184.x
pubmed: 19691645
Prouteau, A. et al. Circulating tumor DNA is detectable in canine histiocytic sarcoma, oral malignant melanoma, and multicentric lymphoma. Sci. Rep. 11, 877 (2001).
doi: 10.1038/s41598-020-80332-y
Beaver, J. A. et al. Detection of cancer DNA in plasma of patients with early-stage breast cancer. Clin. Cancer Res. 20, 2643–2650 (2014).
doi: 10.1158/1078-0432.CCR-13-2933
pubmed: 24504125
pmcid: 4024333
Suppan, C. et al. Sensitive and robust liquid biopsy-based detection of PIK3CA mutations in hormone-receptor-positive metastatic breast cancer patients. Br. J. Cancer 126, 456–463 (2022).
doi: 10.1038/s41416-021-01601-9
pubmed: 34754095
Verret, B., Cortes, J., Bachelot, T., Andre, F. & Arnedos, M. Efficacy of PI3K inhibitors in advanced breast cancer. Ann. Oncol. 30, x12–x20 (2019).
doi: 10.1093/annonc/mdz381
pubmed: 31928690
pmcid: 6923787
André, F. et al. Alpelisib for PIK3CA-mutated, hormone receptor–positive advanced breast cancer. N Engl. J. Med. 380, 1929–1940 (2019).
doi: 10.1056/NEJMoa1813904
pubmed: 31091374
Kim, K. K. et al. Whole-exome and whole-transcriptome sequencing of canine mammary gland tumors. Sci. Data. 6, 147 (2019).
doi: 10.1038/s41597-019-0149-8
pubmed: 31413331
pmcid: 6694171
Goldschmidt, M., Peña, L., Rasotto, R. & Zappulli, V. Classification and grading of canine mammary tumors. Vet. Pathol. 48, 117–131 (2011).
doi: 10.1177/0300985810393258
pubmed: 21266722
Rasotto, R., Berlato, D., Goldschmidt, M. H. & Zappulli, V. Prognostic significance of canine mammary tumor histologic subtypes: An observational cohort study of 229 cases. Vet. Pathol. 54, 571–578 (2017).
doi: 10.1177/0300985817698208
pubmed: 28355108
Peña, L., Andrés, P. D., Clemente, M., Cuesta, P. & Pérez-Alenza, M. Prognostic value of histological grading in noninflammatory canine mammary carcinomas in a prospective study with two-year follow-up: Relationship with clinical and histological characteristics. Vet. Pathol. 50, 94–105 (2013).
doi: 10.1177/0300985812447830
pubmed: 22688585