A novel treatment to enhance survival for end stage triple negative breast cancer using repurposed veterinary anthelmintics combined with gut­supporting/immune enhancing molecules.

Patients with end­stage metastatic disease have limited treatment options and those diagnosed with triple negative breast cancer (Her2, Estrogen receptor, Progesterone receptor) have a poor prognosis. Using a triple negative mammary tumor model selected for brain metastasis (4T1Br4) in the mouse, treatment options that may increase survival when therapeutics are applied at post­metastasis were assessed. Anti­parasitic benzimidazoles (BZs) destabilize microtubules, inhibit metabolic pathways, reduce cell proliferation, and induce apoptosis in tumor cells. Co­administration of two BZs was selected, oxfendazole (OFZ) and parbendazole (PBZ), shown to overcome resistance development in anthelmintic effects by imposing metabolic delay to assess if multiple BZ approach is also suitable to enhance anticancer effects. It has been previously reported that treatment of mammary tumor­bearing mice at an early stage with chitin microparticles (CMPs) decreased tumor growth and metastases by enhancing both innate M1 macrophage and TH1 adaptive immune response. Oral administration of CMPs was previously revealed to affect the gut in intestinal inflammation. A combination BZ (OFZ/PBZ) and CMP treatment was tested to target tumor development and metastasis and effects were compared in response to monotherapies of the same compounds or to untreated mice. The results demonstrated increased survival, decreased tumor cell proliferation, decreased metastasis in lungs and brain, increased levels of fecal SCFAs butyric, acetic, propionic and valeric acids with increased butyric and propionic acid levels in brain biopsies in combination treated compared with untreated mice. At the primary tumor, SCFA receptor FFAR2 expression was increased in combination treatment compared with untreated mice, suggestive of a non­invasive cancer phenotype. The superior cytotoxic effects of OFZ/PBZ were confirmed as opposed to single treatment with OFZ or PBZ using 3D spheroids generated from a human breast cancer cell line, MDA­MB­468. These data are compelling for treatment option possibility even at late stages of metastasized breast cancer.

Progressive generalized tonic-clonic seizures in a transgenic mouse model of adult-onset epilepsy: Implications for morphological changes in cortico-limbic and brainstem structures.

Video/cortical electroencephalography (EEG) is monitored to assess progressive severity of generalized tonic clonic seizures (GTCSs) in a transgenic mouse model of adult-onset epilepsy with increased death risk. The mice overexpress the brain derived neurotrophic factor (BDNF) in the forebrain under the calcium/calmodulin dependent protein kinase 2a (termed TgBDNF) and develop GTCSs in response to tail suspension/cage agitation stimulation at 3-4 months of age. With successive GTCSs (a total of 16 across 10 weeks of assessment), seizures became more severe as evidenced by increased duration of postictal generalized EEG suppression (PGES) associated with loss of posture/consciousness. Mice also developed spike wave discharges with behavioral arrest during the seizure recovery that increased in duration as a function of number of GTCSs. Overall seizure duration (from preictal spike to offset of PGES) and ictal spectral power (full spectra) were also increased. Half of the TgBDNF mice expired following a long period of PGES at the last recorded GTCS. Seizure-evoked general arousal impairment was associated with a striking decrease in total number of gigantocellular neurons of the brainstem nucleus pontis oralis along with increase in volumes of the anterior cingulate cortex and dorsal dentate gyrus in severely convulsive TgBDNF mice compared to litter-matched WT controls and non-convulsive TgBDNF mice. The latter effect was accompanied with an increase in total number of hippocampal granule neurons. These results provide structure-function associations in an animal model of adult-onset GTCSs that progressively increase in severity with clinical relevance for sudden unexpected death following generalized seizures.