In tumors, glucose consumption is elevated in a process known as the Warburg effect and promotes cell growth through an array of mechanisms. Targeting the Warburg effect in tumor cells could be beneficial for treating cancer. Hexokinases phosphorylate glucose in the first rate-limiting step of glycolysis. The hexokinase isoform Hexokinase 1 (HK1) is likely required in most adult mammalian tissues. The hexokinase isoform Hexokinase 2 (HK2) displays increased activity in and is required for the growth of many cancers but global deletion of this isoform is tolerable in mice. A selective HK2 inhibitor would be a promising avenue as a therapeutic. Previous work in our lab identified via high-throughput screening four potential small molecule HK2 inhibitors (Compounds 1 – 4). Our goal was to express recombinant HK2 and characterize the mechanisms behind which HK2 inhibitors exert their target effects. Using a His-tagged bacterial expression vector, we transformed E. coli and induced expression of HK2 via IPTG before extracting the HK2. Compounds 2 – 4 were tested (1) in enzymatic assays against HK1 and HK2 in vitro with varying concentrations of ATP and glucose to assess what type of inhibitions they exerted and to assess their selectivity for HK2 over HK1, (2) in cells that express HK2 to generate dose response curves, and (3) in thermal shift curve assays to determine whether they bind directly to HK2. Compound 3 displayed the greatest potency and selectivity for HK2. Future studies should examine how these compounds might inhibit tumor growth in cell culture tumor models.

Duchenne Muscular dystrophy (DMD) is an inherited degenerative muscular disease caused by a mutation in the DMD gene, which encodes the dystrophin protein. Dystrophin plays a vital role in maintaining cell membrane integrity in muscle fibers via the dystrophin-glycoprotein complex, so loss of function mutations in Dystrophin can cause chronic degeneration in muscle cells. Previous studies done by the Spencer Lab have shown that a global knockout of osteopontin (Spp1) reduces fibrosis and increases muscle strength in mice with DMD. We aim to understand how specific sources of Spp1 affect DMD progression. To do this, the Spencer Lab generated osteopontin conditional knockout (cKO) mouse lines using Cre recombinase. The study of the macrophage Spp1 cKO mouse line showed the disappearance of two stromal cell subpopulations (ApoD and Cxcl13 positive stromal cells) involved in adipogenesis in muscle tissue. Because of the disappearance of these two subpopulations, we hypothesized that there would be a reduction of fat droplets in the muscle tissue of macrophage cKO when compared to control mdx mice. This hypothesis was tested by conducting an oil red o staining of diaphragm muscle of both macrophage cKO and control mdx mice at three months and six months. The results were then subsequently quantified. Although the sample size of this experiment was small, there is a trend of reduced fat droplets in Mϕ cKO diaphragm muscle.

Perinatal mood and anxiety disorders (PMADs) affect 20-25% of birthgivers, and can interfere with pregnancy, delivery and the postpartum period. The 9-item Patient Health Questionnaire (PHQ-9) is used to screen for depression, but may not be ideal for PMAD screening. The 10-item Edinburgh Postnatal Depression Scale (EPDS-10) was developed specifically to screen for PMADs, but is usually delivered on paper or orally. As part of a Postpartum Depression Screening Quality Improvement Initiative, Cedars-Sinai replaced the PHQ-9 with the EPDS-10 and implemented iPads for screening. The self-administrated EPDS-10 will result in more accurate PMAD screen positives and social work referral rates as compared to the nurse-administered PHQ-9. It will also result in fewer incomplete screenings. Through retrospective medical record review, PMAD screen positives and referral rates were identified. Screening positive for depression risk was determined by endorsement of suicidal ideation and/or a PHQ-9 total score of ≥5 or EPDS-10 ≥8. Data were compared before and after the interventions on February 7, 2022. In total, 20,278 individuals delivered a live birth at Cedars-Sinai between 08/31/2020 to 07/31/2023. We expect PMAD screen positive rates closer to epidemiological data (20%), increased social work referral rates, and fewer incomplete screens. Screening with the most appropriate tool improves referral to resources and treatment, thus reducing the risk for adverse mental and physical health outcomes for the birthgiver, child, and family as a whole.