Establishment and characterization of canine mammary tumoroids for translational research

Abstract

Background Cancer heterogeneity is a main obstacle for the development of effective therapies, as its replication in in vitro preclinical models is challenging. Around 96% of developed drugs are estimated to fail from discovery to the clinical trial phase probably because of the unsuitability and unreliability of current preclinical models (Front Pharmacol 9:6, 2018; Nat Rev Cancer 8: 147–56, 2008) in replicating the overall biology of tumors, for instance the tumor microenvironment. Breast cancer is the most frequent cancer among women causing the greatest number of cancer-related deaths. Breast cancer can typically be modeled in vitro through the use of tumoroids; however, current approaches using mouse tumoroids fail to reproduce crucial aspect of human breast cancer, while access to human cells is limited and the focus of ethical concerns. New models of breast cancer, such as companion dogs, have emerged given the resemblance of developed spontaneous mammary tumors to human breast cancer in many clinical and molecular aspects; however, they have so far failed to replicate the tumor microenvironment. The present work aimed at developing a robust canine mammary tumor model in the form of tumoroids which recapitulate the tumor diversity and heterogeneity. Results We conducted a complete characterization of canine mammary tumoroids through histologic, molecular, and proteomic analysis, demonstrating their strong similarity to the primary tumor. We demonstrated that these tumoroids can be used as a drug screening model. In fact, we showed that paclitaxel, a human chemotherapeutic, could kill canine tumoroids with the same efficacy as human tumoroids with 0.1 to 1 μM of drug needed to kill 50% of the cells. Due to easy tissue availability, canine tumoroids can be produced at larger scale and cryopreserved to constitute a biobank. We have demonstrated that cryopreserved tumoroids keep the same histologic and molecular features (ER, PR, and HER2 expression) as fresh tumoroids. Furthermore, two cryopreservation techniques were compared from a proteomic point of view which showed that tumoroids made from frozen material allowed to maintain the same molecular diversity as from freshly dissociated tumor. Conclusions These findings revealed that canine mammary tumoroids can be easily generated and may provide an adequate and more reliable preclinical model to investigate tumorigenesis mechanisms and develop new treatments for both veterinary and human medicine.

Mammary Glands of Women, Female Dogs and Female Rats: Similarities and Differences to Be Considered in Breast Cancer Research

“…By using advanced technologies to manipulate cells, and at different length scales (i.e., 3D/4D bioprinting, mold casting, microfluidic technologies etc. ), multiple replicates of the same tumor can be obtained for various assays, including drug screening [ 215 , 216 , 217 ]. In addition, animals can develop multiple tumors with different histopathological features and molecular subtypes.…”

Cytotoxic effects of crotoxin from Crotalus durissus terrificus snake in canine mammary tumor cell lines

“…Canine cancers occur spontaneously and have a similar clinical presentation and pathophysiology to humans, in addition to sharing the main risk factors and signaling pathways [ 4 , 5 ], such as the expression of the HER-2 protein and Ki-67, related to tumor aggressiveness. This makes dogs a valuable study model as they are more similar to the natural progression of human cancer than induced animal cancer models [ 6 ].…”

Estratégias De Tratamento Do Transtorno Depressivo Recorrente Em Gestante: Um Relato De Caso

“…O câncer de mama é a neoplasia mais frequente em mulheres, causando grande número de mortes devido à doença. Novos modelos de câncer de mama, tais como cadelas, tem surgido dado a semelhança em muitos aspectos clínicos e moleculares de tumores mamários que se desenvolvem espontaneamente nestes animais com o câncer de mama em humanos (RAFFO-ROMERO et al, 2023).…”