Establishment of a bioluminescent canine B-cell lymphoma xenograft model for monitoring tumor progression and treatment response in preclinical studies

Abstract

Canine diffuse large B-cell lymphoma (DLBCL) is one of the most common cancers in dogs which shares remarkable similarities with its human counterpart, making the dog an excellent model for the investigation of novel therapeutic agents. However, the integration of canine lymphoma in comparative studies has been limited due in part to the lack of suitable xenograft mouse models for preclinical studies. To overcome these limitations, we established and characterized a localized subcutaneous bioluminescent canine DLBCL xenograft mouse model. The canine CLBL-1 cell line stably expressing the luciferase and green fluorescent protein reporters was generated and used to establish the xenograft tumor model. A pilot study was first conducted with three different cell densities (0.1×106, 0.5×106 and 1×106 cells) in SCID mice. All mice presented homogeneous tumor induction within eight days after subcutaneous injection, with a 100% engraftment efficiency and no significant differences were observed among groups. The tumors were highly aggressive and localized at the site of inoculation and reproduced histological features and immunophenotype consistent with canine DLBCL. Importantly, xenograft tumors were detected and quantified by bioluminescent imaging. To assess response to therapy, a therapeutic study with a histone deacetylase inhibitor, panobinostat, was performed. The results demonstrated that panobinostat (20 mg/kg) efficiently inhibited tumor growth and that bioluminescent imaging allowed the monitorization and quantification of tumor response to therapy. In summary, this study provides a bioluminescence canine DLBCL model that offers high engraftment efficiency, preservation of tumor features, and noninvasive monitoring of tumor progression, validating the model as a promising preclinical tool for both veterinary and human medicine.

Rabbit derived VL single-domains as promising scaffolds to generate antibody–drug conjugates

“…3 A). The output phages recovered from the 3rd in vitro panning was recovered, reamplified and tailed vein injected in a xenograft CLBL-1 murine model as previously described 39 . The phages were recovered to retrieve sdAbs that specifically targeted and were uptaken by the grafted cNHL tumors.…”

“…The phage library displaying VL sdAbs was first panned using a subtractive cell phage display protocol as previously described by Carlos Barbas 37 , 38 and our studies 25 , and that included a negative selection on HEK293T cells followed by a positive selection on CLBL-1 cells. Then, after three rounds of in vitro selections, an additional panning was performed in vivo in a xenograft CLBL-1 murine model 39 . All animal-handling procedures were performed according to EU recommendations for good practices and animal welfare and approved by the Animal Care and Ethical Committee of the Veterinary Medicine Faculty (Protocol_0050132016).…”

“…The study was conducted according ARRIVE guidelines. Briefly, female 6–8-wk-old SOPF/SHO SCID mice (Charles River) were maintained as mentioned before 39 . Then, the tumors were induced as established previously 39 .…”

Immunotherapeutic Strategies for Canine Lymphoma: Changing the Odds Against Non-Hodgkin Lymphoma

“…However, the implementation of such canine clinical trials is far from being an easy quest. It requires multiple organized efforts to validate the canine model, which still lacks a thorough characterization of the canine immune system and its effector cells and molecules, the evaluation of common tumor epitopes, the development of canine-specific/cross-reactive agents and the establishment of preclinical models for veterinary oncological settings ( 62 , 153 , 154 ). Furthermore, this also requires veterinary scientific community to join forces to implement diagnosis, staging and treatment response assessment optimization and standardization, to perform large and organized clinical trials and to achieve conformity when analyzing data ( 26 ).…”

Epigenetic Mechanisms in Canine Cancer

“…Furthermore, Panobinostat, the most potent HDACi tested in vitro, inhibited CLBL-1 xenograft tumor growth, triggering acetylation of H3 and apoptosis in vivo (204). Panobinostat also efficiently inhibited the growth of tumors in xenograft models inoculated with a modified and bioluminescent canine B-cell lymphoma cell line (205). Trichostatin A (TSA), an antifugical agent with properties to selectively inhibit histone deacetylase activity in mammalian cells has shown inhibitory effects of proliferation and apoptosis in cancer cells (206).…”