Abstract 3382: Detection of cancer in dogs using a novel genetic-based synthetic biomarker platform
Authors: Nieu Regina , Phenix Emily , Hauss Jennifer , Hanson Hadley , Ho Nga , Harwig Alex , Rudina Shireen , Ananthanarayanan Badri , Chang Hong , George Bijee , Louie Maggie C. , Bulman-Fleming Julie , Kent Michael , Suhy David 
Published: June 2022

Abstract
Early detection of cancer is an important driver of increased survival, quality of life and reduced healthcare costs. Although liquid biopsy provides a new option for early cancer detection, the limitations of detecting a few molecules of blood-based biomarkers naturally shed by the cancer remain. Earli is developing a highly sensitive, orthogonal approach that uses a genetic construct that usurps dysregulated pathways and actively forces cancer cells to drive the expression of a detectable ‘synthetic’ biomarker. We previously used murine xenograft models to validate the tumor detection properties of EARLI-001, a circular DNA containing a human derived cancer-activated promoter to drive the transient expression of secreted embryonic alkaline phosphatase (SEAP), a protein normally expressed only during fetal development. Biodistribution following intravenous (IV) administration of EARLI-001 shows a broad tissue tropism, enabling the potential to monitor multiple tissues for malignant cells. A single dose of EARLI-001 administered in a pulmonary metastatic cancer mouse model resulted in a 95-fold increase in serum SEAP compared to healthy controls.
Recognizing the inherent limitations of murine models, we evaluated EARLI-001 in canines diagnosed with cancer. Dog cancers share many features with human malignancies including spontaneous tumor formation, heterogeneity, growth kinetics, histology, and comparable dysregulated genetic pathways. Furthermore, the use of dogs allows the study of EARLI-001 in an immune competent setting and with substantially increased size (up to 65 kg) comparable to scaling into human subjects.
EARLI-001 was first tested for safety and biodistribution in 54 purpose-bred beagles in MTD and GLP toxicology studies which reveal that it is safe and well tolerated across a broad range of clinical doses. Biodistribution analysis in 12 tissues showed nanoplasmid levels peaking in the first days post dose and rapidly decreased to background levels by 60 days. These non-tumor bearing dogs showed no serum SEAP signal at all doses. Following IACUC approval and informed owner consent, EARLI-001 was administered to companion dogs in a standard 3 x 3 dose escalation study, with an expanded mid-range cohort. These companion dogs harbored a wide variety of spontaneous tumors in different tissues of origin. In tumor bearing dogs, EARLI-001 elicited a clear and discernable cancer-activated SEAP signal in 1 of 3 dogs dosed at 0.01 mg/kg, 3 of 6 dosed at 0.03 mg/kg, 2 of 2 at 0.07 mg/kg and 8 of 8 dogs dosed at 0.1 mg/kg. Furthermore, the MTD was not reached, and no significant dose-limiting toxicity was observed in these older, disease-burdened animals. These results show the utility of the platform to interrogate tumor biology and detect the presence of malignancies in relevant in vivo models and provided the critical safety and efficacy data towards a first-in-human clinical study in lung cancer initiated in 2021.
Citation Format: Regina Nieu, Emily Phenix, Jennifer Hauss, Hadley Hanson, Nga Ho, Alex Harwig, Shireen Rudina, Badri Ananthanarayanan, Hong Chang, Bijee George, Maggie C. Louie, Julie Bulman-Fleming, Michael Kent, David Suhy. Detection of cancer in dogs using a novel genetic-based synthetic biomarker platform [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3382.