Combo Group Research Articles

Testosterone and resistance training improved physical performance and reduced fatigue in frail older men: 1 year follow-up of a randomized clinicaltrial.

To improve health conditions among hypogonadal men ≥70 years of age using testosterone undecanoate (TU) injections, progressive strength training, and oral supplements of vitamin D, calcium, and protein. This study is a 1-year follow-up of a double-blind RCT lasting 20 weeks, including 148 older men ≥70 years old with low testosterone levels and mobility problems. During 52 weeks, 4 groups received either testosterone therapy (TU) or progressive resistance training (Training), both (Combo), or no intervention (Controls). Physiotherapists supported the training groups until week 20, while these participants continued trained on their own during weeks 21 to 52. The main outcome measure was the 30-s chair stand test. The following numbers of participants completed the trial: 20 (Combo), 20 (Controls), 24 (TU), and 14 (Training). When examining 30-s chair stand test performance within each group at baseline, and at weeks 4, 20 and 52, only the Combo group improved (p = 0.001, Friedman Test). Compared to controls, only the Combo group experienced reduced fatigue and tiredness (p < 0.05). Fifty-two weeks of testosterone supplementation combined with progressive resistance training may enhance physical performance, alleviate fatigue, and had no notable detrimental impacts among males aged ≥70 suffering from mobility issues and testosterone insufficiency.Trial registration - Clinical Trials NCT02873559.

The Effect of Combination of Cone Drill (Zigzag) with Core Stability, Combination of Ladder Drill (Snake Jump) with Core Stability, and Speed on Agility of Futsal Players: A Factorial Experimental Design

The game of futsal, known for its high intensity and dynamic nature demanding skill and robust physical fitness, particularly in agility, presents a significant challenge. Incorporating variations of cone and ladder drills in agility training, with a focus on enhancing balance through core stability training in conjunction with speed modulation as a key factor, is believed to impact players' agility players. Despite recognizing the individual significance of these training modalities in boosting agility performance, there remains a gap in fully comprehending their combined effects during futsal engagements. This research seeks to scrutinize the group differentials in agility training, factoring in speed variations in relation to futsal players' agility levels. The study defined agility training cohorts as a combo group of cone drill with core stability and a combination group of ladder drill alongside core stability, while speed was categorized as high or low. Employing an experimental methodology with a 2 x 2 factorial design, the study enlisted 20 players aged 17.1 ± 0.76 years through random selection. Agility training, conducted thrice weekly over a six-week period, emphasized swift and dynamic movements alongside maintaining balance. Speed and agility metrics were assessed using the 30m acceleration test and Illinois agility test, then subjected to statistical scrutiny via a two-way ANOVA at a significance level of 0.05. Results revealed that both training methodologies benefited participants with elevated speed (P&lt;0.05). Notably, agility training incorporating ladder drill exercises and core stability training exhibited superior outcomes for high-speed performers (P&lt;0.05). Conversely, individuals with lower speed levels potentially benefitted more from a combination of ladder drill and core stability (P&lt;0.05). These findings furnish crucial insights for futsal coaches and trainers looking to craft tailored training regimens aimed at enhancing agility performance effectively. Keywords: Cone Drill, Ladder Drill, Core Stability, Speed, Agility, Futsal

Immuno-related cardiac toxicity in patients with cancer: A prospective study applying multiparametric cardiac MRI.

e24014 Background: Since Immune checkpoint inhibitor (ICI) immunotherapy has emerged as a cornerstone of modern oncology, oncologists are facing a new spectrum of ‘immune-related adverse events’ (irAEs), potentially affecting every tissue and organ. ICI-induced symptomatic myocarditis occurs only in around 1% of ICI-treated patients (pts), but is fatal in 50% of cases. Methods: Pts undergoing ICI therapy at the Humanitas Cancer Center from May 2021 to December 2023 were prospectively recruited and assigned to three study groups: 1) ICI given as monotherapy (Mono) 2) ICI given in combination (Combo) 3) history of cardiac disease or two cardiological risk factors other than smoke (Cardio). Exclusion criteria were: previous exposure to any immunotherapy, combination with other cardiotoxic therapies, and acute (in the last 3 months) cardiac events. All pts underwent a complete cardiological assessment with clinical visit, 12-lead ECG, and multiparametric cardiac MRI (cMRI) at two time-points: prior initiation of the ICI therapy, and around 8 weeks later. Peripheral blood samples for immunophenotyping analyses were collected at the same time-points. cMRI scans were performed using a 1.5 Tesla scanner. Chi-square, Fisher exact and paired t-test were used to compare groups; all analyses were performed using SAS v9.4. Objectives of the study were: detection of subclinical cardiac damage and identification of groups of patients at major risk, who may benefit from cardiological pre- or/and on-treatment assessment. Results: We present data on the first 47 pts enrolled, diagnosed with a wide range of solid tumors, including 16 renal carcinoma, 11 melanoma and 3 NSCLC; 15 (32%), 16 (34%) and 16 (34%) pts were assigned to the Mono, Combo and Cardio group, respectively. Most pts (42, 89%) were treated with ICI for locally advanced/metastatic disease (26 as first-line therapy, 16 from second-line on), while 5 (11%) as adjuvant therapy. All pts received an anti-PD-(L)1, either as single agent (25, 53%) or in combination, with ICI plus a TKI being the most frequent combinatorial strategy (16; 34%). Overall, the cMRI analysis showed a statistically significant left ventricle ejection fraction (LVEF) reduction pre- versus post-ICI treatment (p &lt; 0.001), with 18 (38%) pts experiencing a loss of &gt; 3 points of LVEF. No statistically significant differences in LVEF reduction were observed considering the three different groups (p = 0.6), tumor types (p = 0.2), or occurrence of any grade non-cardiological irAEs (p = 0.6). Conclusions: Our study was able to show a statistically significant reduction in LVEF, with more than 3% LVEF loss in a clinically significant proportion of ICI-treated patients (18; 38%), not selected for suspected cardiac symptoms, warranting prospective evaluation to eventually identify pts at higher risk. The trial is continuing prospective enrollment and immunophenotyping analyses are ongoing.

Therapy Combining Glucagon-Like Peptide-1 Receptor Agonist with Sodium-Glucose Cotransporter 2 Inhibitor Suppresses Atherosclerosis in Diabetic ApoE-Deficient Mice.

Glucagon-like peptide-1 receptor agonists (GLP-1RAs) and sodium-glucose cotransporter 2 inhibitors (SGLT2i) have beneficial effects on cardiovascular disease in addition to their glucose-lowering effects. In this study, the effects of these drugs, when used individually or in combination, on cardiovascular atherosclerotic lesion development were compared in diabetic ApoE-deficient (ApoE KO) hyperlipidemic mice. ApoE-KO mice were treated with streptozotocin and nicotinamide, generating a type 2 diabetes model. The mice were randomly divided into four groups: vehicle-treated (untreated), liraglutide (LIRA), ipragliflozin (IPRA), and combination therapy (combo). These mice, as well as non-diabetic controls, were fed a high-fat diet. After 8 weeks of drug administration, the heart and aorta were removed and analyzed. Atherosclerotic lesions evaluated by oil red O (ORO) staining were significantly larger in the untreated group (13.4±0.8% of the total aortic area) than in the non-diabetic controls (4.4±0.5%, p<0.01), while being reduced in the combo group (6.0±1.0%, p<0.01) as compared with the untreated group. The ORO stain-positive area in the LIRA and IPRA groups tended to be reduced but their differences were not statistically significant. Transcript levels of Mcp1 and Sirt1 were significantly reduced and increased, respectively, in the combo compared with the untreated group, while no significant changes were observed in the monotherapy groups. The data suggest that combination therapy with liraglutide and ipragliflozin may be an efficient regimen for preventing the development of atherosclerosis in diabetic mice deficient in ApoE.

Assessment of the role of false-positive alerts in computer-aided polyp detection for assistance capabilities.

False positives (FPs) pose a significant challenge in the application of artificial intelligence (AI) for polyp detection during colonoscopy. The study aimed to quantitatively evaluate the impact of computer-aided polyp detection (CADe) systems' FPs on endoscopists. The model's FPs were categorized into four gradients: 0-5, 5-10, 10-15, and 15-20 FPs per minute (FPPM). Fifty-six colonoscopy videos were collected for a crossover study involving 10 endoscopists. Polyp missed rate (PMR) was set as primary outcome. Subsequently, to further verify the impact of FPPM on the assistance capability of AI in clinical environments, a secondary analysis was conducted on a prospective randomized controlled trial (RCT) from Renmin Hospital of Wuhan University in China from July 1 to October 15, 2020, with the adenoma detection rate (ADR) as primary outcome. Compared with routine group, CADe reduced PMR when FPPM was less than 5. However, with the continuous increase of FPPM, the beneficial effect of CADe gradually weakens. For secondary analysis of RCT, a total of 956 patients were enrolled. In AI-assisted group, ADR is higher when FPPM≤5 compared with FPPM>5 (CADe group: 27.78% vs 11.90%; P=0.014; odds ratio [OR], 0.351; 95% confidence interval [CI], 0.152-0.812; COMBO group: 38.40% vs 23.46%, P=0.029; OR, 0.427; 95% CI, 0.199-0.916). After AI intervention, ADR increased when FPPM≤5 (27.78% vs 14.76%; P=0.001; OR, 0.399; 95% CI, 0.231-0.690), but no statistically significant difference was found when FPPM>5 (11.90% vs 14.76%, P=0.788; OR, 1.111; 95% CI, 0.514-2.403). The level of FPs of CADe does affect its effectiveness as an aid to endoscopists, with its best effect when FPPM is less than 5.

Abstract 3739: Different combination regimens for OX40 agonists and PD-1 inhibitors exert different antitumor effects in OX40 humanized mice

Abstract OX40, also known as CD134 or TNFRSF4, is a member of the tumor necrosis factor receptor (TNFR) superfamily which mainly expressed on the surface of activated T cells including Treg cells. Engagement of OX40 with its ligand, TNFSF4, leads to effector T cell expansion and cytokine release, inhibits activation-induced cell death (AICD) and promotes the survival of antigen-specific memory T cells. In contrast to the massive hit of immune checkpoint inhibitors like PD-1 and PD-L1, the benefit of agonistic antibodies in clinical trials has not been completely demonstrated to date. On one hand, higher doses could leads to T cell exhaustion or AICD, paradoxically, with no efficacy improvement. Another hand, the limited understanding of the biology of immune checkpoints underlying “accelerator and brake” should account for the slow clinical progress of OX40-targeted biologics. It seems that the dosing sequence or timing of anti-OX40 and anti-PD-1 is an important consideration in combination therapies. Given this, we firstly evaluated the antitumor effects of two different anti-human OX40 agonists in syngeneic MC38 and Pan02 tumor models, respectively, in hOX40 knockin mice. We observed tumor regressions upon treatment of OX40 agonistic Ab1 in a dose-dependence way in the homozygous hOX40 knockin mice engrafted with MC38 tumors. The anti-hOX40 Ab1 at high-dose (10 mg/kg) led to ~60% TGI, but no response at very low dose (0.1 mg/kg). Furthermore, the homozygous hOX40 knockin mice inoculated with Pan02 pancreatic tumors were treated with a novel strong anti-hOX40 agonist (Ab2) and surrogate anti-mPD-1. Notably, Anit-hOX40 (Ab2) alone at low dose (1 mg/kg, Q3D*9 doses) exerted strong anti-tumor potency (TGI=59.3%, p&amp;lt; 0.005) and no statistical significant antitumor difference was observed between high-dose group (at 5mg/kg) and low-dose group (at 1 mpk) as monotherapy, though anti-hOX40 (Ab2) at 5mg/kg showed the best therapeutic response (TGI=72%, p=0.0002) among all treatment groups. In contrast, the second therapeutic response was observed in the combo group treated with first dosing of 5mg/kg anti-hOX40 (Ab2) combining delayed dosing of 10mg/kg anti-mPD-1 (TGI=64%, p&amp;lt; 0.005) and the concurrent dosing group of Ab2 (5 mg/kg, Q3D × 9 doses) plus anti-mPD-1 (2 mg/kg, Q3D × 9 doses) (TGI=62.3%, p= 0.001). In conclusion, different human-specific OX40 agonists displayed different anti-tumor activity as monotherapy, and different dose led to different antitumor potency for the same agonist. In terms of different combination regimens, although no advanced benefit was observed for the combination of anti-mPD-1/anti-OX40 (Ab2) in comparison to strong anti-OX40 agonist (Ab2) alone, it demonstrated that the dosing timing and sequence of either anti-PD-1 or anti-hOX40 did has impact on the therapeutic outcome. Citation Format: Yi Li, Yanlei Zhang, Xiaolei Qiu, Dongxiao Feng, Ruilin Sun, Daniel X. He. Different combination regimens for OX40 agonists and PD-1 inhibitors exert different antitumor effects in OX40 humanized mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 3739.

Infliximab Monotherapy vs Combination Therapy for Pediatric Crohn's Disease Exhibit Similar Pharmacokinetics.

The use of concomitant azathioprine may improve efficacy and pharmacokinetic (PK) properties of infliximab (IFX) but is also associated with an increased risk of adverse events. Proactive therapeutic drug monitoring (pTDM) of IFX monotherapy is an alternative strategy to improve PK. The aim of this study was to evaluate whether IFX with an immunomodulator (combo) has PK benefits over IFX-pTDM (mono) in pediatric Crohn's disease (CD). This PK analysis included pediatric CD patients who started either IFX combo (TISKids study) or IFX mono with pTDM (REFINE cohort). Combo and mono IFX trough levels (TLs) and antibodies-to-infliximab were assessed at infusion 3, 4, and 5. A population PK model was built to compare IFX PK outcomes (clearance [CL], TLs and cumulative exposure) between combo and mono groups at infusion 4 and 5. Clinical response and steroid-free clinical remission (SFCR) was assessed at infusion 4 and 5. This study included 128 pediatric CD patients (66 mono and 62 combo). At infusion 5, there was no significant difference between mono and combo median TLs 4.1 µg/mL (2.1, 7.8) vs 5.9 µg/mL (3.2, 9.4; P = .14) or median CL 0.26 L/d (0.21, 0.32) vs 0.26 L/d (0.21, 0.33; P = .81). Mono patients had a lower SFCR rate at infusion 5 (53% [31 of 59] vs 80% [32 of 40]; P = .01). Clinical response rates were significantly higher among combo than mono patients at both infusion 4 and 5. This study suggests that there are no PK differences (TLs and CL) between combo and mono therapy in pediatric CD patients who started IFX.

Synergistic Growth Inhibition of NPM1 Mutant AML PDX By Combined Therapy with BCL-2 Inhibitor Venetoclax (ABT-199) and Menin Inhibitor DS-1594b In Vivo

Introduction: Mixed-lineage-leukemia (MLL) translocations are prevalent in hematologic malignancies, resulting in aggressive leukemias resistant to standard therapies and low 5-year survival rates. These translocations lead to the production of MLL fusion proteins that promote leukemia development by enhancing proliferation, inhibiting hematopoietic differentiation, and driving leukemogenesis. Acute myeloid leukemia (AML), an aggressive blood cancer with a grim prognosis, particularly in elderly patients, frequently involves mutations in the nucleophosmin (NPM1) gene. Importantly, the interaction between Menin, MLL, and MLL fusion proteins play a crucial role in the pathogenesis of AML, both with MLL rearrangements (MLLr) and in leukemias carrying NPM1 mutations. Inhibition of the Menin-MLL interaction using novel Menin inhibitors has demonstrated efficacy in pre-clinical and clinical studies for both AML subsets. In this study, we investigate the effectiveness of the novel Menin-MLL inhibitor DS-1594b, developed by Daiichi Sankyo Co. Ltd., in combination with the BCL-2 antagonist venetoclax (ABT-199), in a patient-derived xenograft (PDX) model of NPM1-mutated acute leukemia. We have previously reported synergistic lethality and differentiation-inducing effects of this drug combination in both AML cell lines and primary leukemia samples in vitro (Ciaurro et al., Blood 2022). Our findings shed light on the potential of this therapeutic combination for the targeted treatment of AML with NPM1 mutations. Methods: NSG mice (female, 8-10 weeks old) were purchased from Jackson Laboratory. Mice were inoculated via the tail vein with NPM1m PDX/luc/GFP (3×10 6 cells). Once leukemia engraftment was confirmed by bioluminescent imaging mice were randomized in 4 groups: vehicle, ABT199, DS-1594b alone, and combination. After 12 days drugs were administered as follows: DS1594b at a dosage of 50 mg/kg by oral gavage for 4 weeks; ABT-199 at a dosage of 50 mg/kg by oral gavage for 2 weeks and 100mg/kg for the other 2 weeks. NSG mice were euthanized by CO 2 inhalation at the endpoint of the study in accordance with IACUC protocols. BM and spleens were harvested in a sterile environment and flushed in HBSS with 1% FBS. For CyTOF data spleen cells were initially barcoded and stained with metal-conjugated antibodies according to Fluidigm's protocols and samples were analyzed by CyTOF (Helios) Mass Cytometer. Results: New in vitro data confirmed synergistic lethality on additional AML cell lines and demonstrated upregulation of PARP-C and Caspase3-C apoptosis pathways using western blot analysis. Further, additional 3 primary AML patient samples showed additive or synergistic growth inhibition when treated with combination of drugs. In the in vivo experiment with NPM1m PDX/luc/GFP AML PDX model, we found that co-treatment with DS-1594b and venetoclax for four weeks resulted in a greater reduction in AML burden compared to treatment with either agent alone or a vehicle control as shown by Bioluminescence imaging. Moreover, weight of mice in the combo group was not significantly affected, while mice lost weight in vehicle and in single agent cohort due to rapid disease progression. The survival curve clearly demonstrated the benefits of the combination treatment, with the DS-1594b group having modest anti-tumor efficacy, and the combination group showing the greatest efficacy (Fig. 1A). To study the effects on cell differentiation, we used CyTOF technique, which allowed us to analyze a complete panel of 30 markers. We found two populations of cells, which we named populations (pop) 3 and pop 4, that were enriched in the group treated with the combination of drugs compared to the control group (Fig 1B). These populations of cells had higher levels of expression of CD33 and CD38, which are well-known markers of cellular differentiation. Conclusions: In this study, we present the in vivo effectiveness of DS-1594b in combination with venetoclax in a PDX model of NPM1-mutated AML. Moreover, the combination treatment exhibits differentiation-inducing effects, which contribute to its therapeutic effectiveness, and was safe. Overall, our findings strongly indicate that the combination of DS-1594b and venetoclax exhibits promising anti-leukemic activity in vivo and holds potential as a therapeutic strategy for AML patients with mtNPM1 mutations.

Expression of EGFR-mutant proteins and genomic evolution in EGFR-mutant transformed small cell lung cancer.

The transformation of epidermal growth factor receptor (EGFR)-mutant lung adenocarcinoma (LUAD) into small cell lung cancer (SCLC) accounts for 3-14% of the resistance mechanism to EGFR tyrosine kinase inhibitors (TKIs). At present, there is no relevant research to explore the dynamic expression of EGFR-mutant proteins and genomic evolution in EGFR-mutant transformed SCLC/neuroendocrine carcinoma (NEC). Genetic analysis and protein level analysis by next-generation sequencing (NGS), Whole-exome sequencing (WES) and immunohistochemistry were performed to explore expression of EGFR-mutant proteins and genomic evolution in EGFR-mutant transformed SCLC. The research used three patient-derived organoids (PDOs) to explore the efficacy of combo [chemotherapy (chemo) plus TKI or bevacizumab] treatment. According to the subsequent treatment regimens after SCLC/NEC transformation, 35 patients were divided into chemo (n=21) and combo (n=14) groups. EGFR L858R and EGFR E746-750 del protein expression by immunohistochemistry was 80.0% (4/5) and 100% (6/6), respectively (P=0.455) in initially-transformed tissues. Meanwhile, EGFR-mutant proteins were expressed in 85.7% (6/7) of dynamic rebiopsy tissues or effusion samples after the first transformation. Then, by the pathway enrichment analysis of tissue and plasma NGS, the EGFR-related pathways were still activated after SCLC/NEC transformation. Moreover, WES analysis revealed that transformed SCLC shared a common clonal origin from the baseline LUAD. The drug sensitivity of three PDOs demonstrated potent anti-cancer activity of EGFR-TKIs plus chemo, compared with chemo or TKI alone. There were significant differences in objective response rate (ORR) between the combo and chemo groups [42.9 % vs. 4.8%, P=0.010, 95% confidence interval (CI): 1.5-145.2]. Furthermore, the median post-transformation progression-free survival (pPFS) was significantly prolonged in the combo group, with 5.4 (95% CI: 3.4-7.4) versus 3.5 (95% CI: 2.7-4.3, P=0.012) months. EGFR 19del or L858R-mutant proteins could be constantly expressed, and EGFR pathway still existed in EGFR-mutant transformed SCLC/NEC with a common clonal origin from the baseline LUAD. Taking together, these molecular characteristics potentially favored clinical efficacy in transformed SCLC/NEC treated with the combo regimen.

Pressure-Enabled Drug Delivery (PEDD) of a class C TLR9 agonist in combination with checkpoint inhibitor therapy in a murine pancreatic cancer model

BackgroundSystemic immunotherapy has had limited clinical benefit in pancreatic ductal adenocarcinoma. This is thought to be due to its desmoplastic immunosuppressive tumor microenvironment in addition to high intratumoral pressures that limit drug delivery. Recent preclinical cancer models and early-phase clinical trials have demonstrated the potential of toll-like receptor 9 agonists, including the synthetic CpG oligonucleotide SD-101, to stimulate a wide range of immune cells and eliminate suppressive myeloid cells. We hypothesized that Pressure-Enabled Drug Delivery via Pancreatic Retrograde Venous Infusion of toll-like receptor 9 agonist would improve responsiveness to systemic anti-programmed death receptor-1 checkpoint inhibitor therapy in a murine orthotopic pancreatic ductal adenocarcinoma model. MethodsMurine pancreatic ductal adenocarcinoma (KPC4580P) tumors were implanted into the pancreatic tails of C57BL/6J mice and treated 8 days after implantation. Mice were assigned to one of the following treatment groups: Pancreatic Retrograde Venous Infusion delivery of saline, Pancreatic Retrograde Venous Infusion delivery of toll-like receptor 9 agonist, systemic anti-programmed death receptor-1, systemic toll-like receptor 9 agonist, or the combination of Pancreatic Retrograde Venous Infusion delivery of toll-like receptor 9 agonist and systemic anti-programmed death receptor-1 (Combo). Fluorescently labeled toll-like receptor 9 agonist (radiant efficiency) was used to measure uptake of the drug on day 1. Changes in tumor burden were evaluated by necropsy at 2 different time points, 7 and 10 days after toll-like receptor 9 agonist treatment. Blood and tumors were collected at necropsy 10 days after toll-like receptor 9 agonist treatment for flow cytometric analysis of tumor-infiltrating leukocytes and plasma cytokines. ResultsAll mice analyzed survived to necropsy. Site of tumor fluorescence measurements revealed 3-fold higher intensity fluorescence in Pancreatic Retrograde Venous Infusion delivery of toll-like receptor 9 agonist compared to systemic toll-like receptor 9 agonist mice. Tumor weights were significantly lower in the Combo group compared to Pancreatic Retrograde Venous Infusion delivery of saline. Flow cytometry of the Combo group demonstrated significantly increased overall T-cell number, specifically CD4+ T-cells, and a trend toward increased CD8+ T-cells. Cytokine analysis showed significantly decreased IL-6 and CXCL1. ConclusionPressure-Enabled Drug Delivery of toll-like receptor 9 agonist by Pancreatic Retrograde Venous Infusion with systemic anti-programmed death receptor-1 demonstrated improved pancreatic ductal adenocarcinoma tumor control in a murine pancreatic ductal adenocarcinoma model. These results support study of this combination therapy in pancreatic ductal adenocarcinoma patients and expansion of ongoing Pressure-Enabled Drug Delivery clinical trials.

1813-PUB: Cardiorenal Effect of Dapagliflozin and Dapagliflozin-Saxagliptin Combination on CD34+ve Hematopoietic Stem Cells (HSCs) and Podocyte Specific Markers in Type 2 Diabetes Subjects

Introduction: Effect of Dapagliflozin (Dapa) and Dapagliflozin-Saxagliptin combination (Combo) was examined on peripheral blood derived CD34+ HSCs as a cellular CVD biomarker. Both Dapa (a SGLT2 receptor inhibitor) and Saxagliptin (a DPP4 enzyme inhibitor) are T2DM meds, but the combo has not been evaluated for cardio-renal risk. Hypothesis: We hypothesized that Dapa will improve the outcomes when compared to placebo and the Combo maybe even more beneficial. Methods: 15 subjects were enrolled in 16 weeks, double-blind, three-arm, randomized placebo matched trial, with 10mg Dapa + Saxa placebo (n=4), 10 mg Dapa + 5 mg Saxa (n=5) Combo, And Dapa placebo + Saxa placebo (n=6), Placebo groups. T2DM (30-70 yrs) with HbA1c of 7-10%, were included. CD34+ HSC number, migration, mRNA expression along with biochemistry and urine exosomes were measured. Data were collected at wks 0, 8, and 16. For stats, a mixed model regression analysis was used. Results: Significant HbA1c (p=0.0357) reduction was noted in Combo group vs Dapa alone and Placebo. hsCRP levels (P=0.0317) and IL-6, two important inflammatory molecules, were significantly reduced in both Dapa and Combo vs Placebo. Leptin levels decreased significantly in both Dapa alone (p=0.035) and Combo group (p=0.015), vs Placebo, however the Adiponectin levels were higher in Dapa alone group. EPC migration improved significantly in both Dapa alone (p=0.05) and Combo group (p=0.05) vs Placebo. Interestingly, urine exosome-based podocyte specific protein such as podocalyxin was actually higher in Combo vs Control. Conclusions: Several parameters showed significant improvement with both Dapa and Combo vs placebo. However, other than glycemic control Combo didn’t offer any further benefit over Dapa alone in terms of HSC and biochemistry. Moreover, the Combo may have deleterious effect on renal podocyte inflammation which needs to be examined in a larger study. Disclosure S.Nandula: None. S.Sen: None.

RAMP 202: A phase 2 study of avutometinib (VS-6766) ± defactinib, in patients with advanced KRAS G12V mutant non–small cell lung cancer (NSCLC).

9100 Background: KRAS mutations (mt) occur in ~30% of lung adenocarcinomas, among which G12C is most common (40%), followed by G12V (22%) and G12D (16%). Approved treatments for advanced KRAS mt NSCLC (excluding G12C) are limited to chemotherapy and immune checkpoint inhibitors (ICIs). Avutometinib is a novel small molecule RAF/MEK clamp. Focal adhesion kinase (FAK) activation is a resistance mechanism to RAF/MEK inhibition. Defactinib, a small molecule FAK inhibitor, has shown synergistic antitumor activity with avutometinib in preclinical models. In prior studies, avutometinib ± defactinib has shown responses in patients (pts) with KRAS mt NSCLC, including KRAS G12V. Methods: RAMP 202 is a randomized, phase 2, adaptive, multicenter, open-label study evaluating the efficacy and safety of avutometinib ± defactinib in previously-treated KRAS mt NSCLC (NCT04620330). Key inclusion criteria include histologically confirmed NSCLC with known KRAS mt and ≥1 prior systemic therapy (platinum-based and immune checkpoint inhibitor or appropriate therapy for activating mutation). Part A evaluated the optimal regimen, either 4.0 mg avutometinib orally (PO), twice weekly, 3 weeks on, 1 week off (mono) or 3.2 mg avutometinib PO twice weekly + 200 mg defactinib PO twice per day, 3 weeks on, 1 week off (combo) in pts with KRAS G12V mt NSCLC. Primary endpoint was confirmed objective response rate (ORR) by blinded independent central review. The optimal regimen determined in Part A would subsequently be assessed for efficacy in Part B. Exploratory assessment of ORR is also planned in KRAS-other (non-G12V) NSCLC. Results: Of 35 pts with KRAS G12V enrolled to Part A, 16 pts received mono, and 19 received combo. Patients received up to 5 lines of prior systemic therapy (median 2), including prior platinum-based chemotherapy, ICIs, and bevacizumab. No confirmed responses were seen in the mono group. In the combo group, 2 pts (11%) experienced a confirmed partial response. The duration of each response was 7.9 and 8.5 months, with both ongoing at data cut. The majority of treatment-related adverse events (TRAEs, any grade) (N = 72) were mild to moderate. The most common Grade ≥3 TRAEs included blood CPK increase (11.1%), diarrhea (5.6%), anemia (5.6%), and rash (1.4%). Most AEs were manageable/reversible. Conclusions: In this heavily pretreated population of patients with KRAS G12V mt NSCLC, limited clinical activity was observed with combination therapy. While no new safety signals were identified, criteria to proceed to part B were not met, and further evaluation of avutometinib ± defactinib in KRAS G12V mt NSCLC will not be pursued. Additional trials evaluating rational avutometinib combinations (sotorasib, adagrasib, everolimus) are ongoing in patients with KRAS mt NSCLC. Clinical trial information: NCT04620330 .

A CXCR4 inhibitor (balixafortide) enhances docetaxel-mediated antitumor activity in a murine model of prostate cancer bone metastasis.

Prostate cancer (PCa) bone metastases have been shown to be more resistant to docetaxel than soft tissue metastases. The proinflammatory chemokine receptor CXCR4 has been shown to confer resistance to docetaxel (DOC) in PCa cells. Balixafortide (BLX) is a protein epitope mimetic inhibitor of CXCR4. Accordingly, we hypothesized that BLX would enhance DOC-mediated antitumor activity in PCa bone metastases. PC-3 luciferase-labeled cells were injected into the tibia of mice to model bone metastases. Four treatment groups were created: vehicle, DOC (5 mg/kg), BLX (20 mg/kg), and combo (receiving both DOC and BLX). Mice were injected twice daily subcutaneously with either vehicle or BLX starting on Day 1 and weekly intraperitoneally with DOC starting on Day 1. Tumor burden was measured weekly via bioluminescent imaging. At end of study (29 days), radiographs were taken of the tibiae and blood was collected. Serum levels of TRAcP, IL-2, and IFNγ levels were measured using ELISA. Harvested tibiae were decalcified and stained for Ki67, cleaved caspase-3, and CD34 positive cells or microvessels were quantified. Tumor burden was lower in the combo group compared to the DOC alone group. Treatment with the combination had no impact on the number of mice with osteolytic lesions, however the area of osteolytic lesions was lower in the combo group compared to the vehicle and BLX groups, but not the DOC group. Serum TRAcP levels were lower in the combo compared to vehicle group, but not the other groups. No significant difference in Ki67 staining was found among the groups; whereas, cleaved caspase-3 staining was lowest in the Combo group and highest in the BLX group. The DOC and combo groups had more CD34+ microvessels than the control and BLX groups. There was no difference between the treatment groups for IL-2, but the combo group had increased levels of IFNγ compared to the DOC group. Our data demonstrate that a combination of BAL and DOC has greater antitumor activity in a model of PCa bone metastases than either drug alone. These data support further evaluation of this combination in metastatic PCa.

505 Risk Stratification by TBSA for Acute Kidney Injury in Burn Patients Following Combination Antibiotic Therapy

Abstract Introduction Burn patients are susceptible to infections and are often treated with broad-spectrum antibiotics. Vancomycin is a common antibiotic administered alone, or in combination with piperacillin-tazobactam (Pip/Tazo). This combination therapy was associated with increased rates of acute kidney injury (AKI) in burn patients when compared to Vancomycin alone or in combination with other antibiotics. However, recent studies failed to demonstrate these findings. We sought to determine the degree of nephrotoxicity of Vancomycin, Pip/Tazo, and their combination, with burn severity. Methods A retrospective, multi-institutional study was performed utilizing TriNetX Global Health Research Network that encompasses de-identified patient records from 55 health care organization across the US. We identified the population of burn patients between 2000-2020 and stratified them into 3 groups by TBSA &amp;lt; 10%, 10-39%, or 40%+. Patients were further stratified into cohorts by administration of broad-spectrum antibiotics within 3 months of their burn: Vancomycin, Pip/Tazo alone, in combination (Combo), or neither. Cohorts were propensity score matched for age at index, gender, race, ethnicity, and pre-existing conditions that predispose to AKI. We explored patient distribution and conducted a comparative outcome analysis to determine the risk of developing AKI in 7- and 90-day outcome windows. Results Vancomycin and Pip/Tazo were compared to control groups and showed increased risk of AKI in all TBSA groups and time outcomes(p&amp;lt; 0.0001). Before stratifying by TBSA, all burn patients administered combination antibiotics showed increased risk of AKI when compared to control, Vancomycin alone, and Pip/Tazo. In patients under &amp;lt; 10% TBSA, combo antibiotics increased AKI incidence in both 7- and 90-days when compared to all other groups. In the 10-39% TBSA group, combination therapy increased the risk of AKI when compared to Pip/Tazo at 7-days. Increased risk of AKI occurred in the combo group when compared to both Vancomycin and Pip/Tazo in 90-days. In the 40%+ TBSA cohort, combination antibiotics did not increase the incidence of AKI in either time period or when compared to Vancomycin and Pip/Tazo. The results of the comparative analysis are included in Table 1. Conclusions Broad-spectrum antibiotics use in burn patients is associated with increased risk of AKI, this risk is higher with Vancomycin and Piperacillin/Tazobactam combination. Combo therapy increased AKI in &amp;lt; 10% and 10-39% but not 40%+ TBSA. These findings can be used to monitor for early manifestations of AKI when using broad-spectrum antibiotics. Applicability of Research to Practice The decision to initiate antimicrobial therapy is crucial in burn patients as they are already at risk for AKI. TBSA can be a useful tool for guiding risk stratification.

Dual Inhibition of the Wnt Inhibitors DKK1 and Sclerostin Promotes Fracture Healing and Increases the Density and Strength of Uninjured BoneAn Experimental Study in Nonhuman Primates.

Fracture repair involves the reactivation of developmental signaling cascades, including Wnt signaling that stimulates bone formation and bone regeneration. Rodent data indicate that dual inhibition of the Wnt signaling antagonists sclerostin and Dickkopf-1 (DKK1) increases callus bone volume and strength while increasing bone mass systemically. We evaluated the effects of 16 weeks of subcutaneously administered carrier solution (vehicle, VEH), anti-sclerostin antibody (Scl-Ab), anti-DKK1 antibody (DKK1-Ab), or Scl-Ab plus DKK1-Ab combination therapy (COMBO) on ulnar osteotomy healing in nonhuman primates (cynomolgus monkeys; 20 to 22 per group). Scl-Ab and COMBO therapy increased systemic markers of bone formation versus VEH, with COMBO leading to synergistic increases versus Scl-Ab or DKK1-Ab monotherapies. The COMBO and Scl-Ab groups showed reduced serum markers of bone resorption versus VEH. The COMBO and DKK1-Ab groups exhibited greater callus bone mineral density (BMD), torsional stiffness, and torsional rigidity versus VEH. Lumbar vertebrae from the Scl-Ab and COMBO groups showed greater BMD and bone formation rate versus VEH, and the femoral mid-diaphysis of the Scl-Ab and COMBO groups showed greater periosteal and endocortical bone formation rates versus VEH. DKK1-Ab increased BMD and strength at the ulnar osteotomy site, Scl-Ab increased bone formation and BMD at uninjured skeletal sites, and Scl-Ab plus DKK1-Ab combination therapy induced all of these effects, in some cases to a greater degree versus 1 or both monotherapies. These results in nonhuman primates suggest that DKK1 preferentially regulates bone healing while sclerostin preferentially regulates systemic bone mass. Combination therapy with antibodies against sclerostin and DKK1 may offer a promising therapeutic strategy for both fracture treatment and fracture prevention.

Abstract 691: Cardio-renal Effect Of Dapagliflozin Alone Versus Dapagliflozin- Saxagliptin Combination In Type 2 Diabetes (t2dm) Subjects

Introduction: Effect of Dapagliflozin (Dapa) and Dapagliflozin-Saxagliptin combination (Combo) was examined on peripheral blood derived CD34+ HSCs as a cellular CVD biomarker. Both Dapa (a SGLT2 receptor inhibitor) and Saxagliptin (a DPP4 enzyme inhibitor) are T2DM meds, but the combo has not been evaluated for cardio-renal risk. Hypothesis: We hypothesized that Dapa will improve the outcomes when compared to placebo and the Combo maybe even more beneficial. Methods: 15 subjects were enrolled in 16 weeks, double-blind, three-arm, randomized placebo matched trial, with 10mg Dapa + Saxa placebo (n=4), 10 mg Dapa + 5 mg Saxa (n=5) Combo, And Dapa placebo + Saxa placebo (n=6), Placebo groups. T2DM (30-70 yrs) with HbA1c of 7-10%, were included. CD34+ HSC number, migration, mRNA expression along with biochemistry and urine exosomes were measured. Data were collected at wks 0, 8, and 16. For stats, a mixed model regression analysis was used. Results: Significant HbA1c (p=0.0357) reduction was noted in Combo group vs Dapa alone and Placebo. hsCRP levels (P=0.0317) and IL-6, two important inflammatory molecules, were significantly reduced in both Dapa and Combo vs Placebo. Leptin levels decreased significantly in both Dapa alone (p=0.035) and Combo group(p=0.015), vs Placebo, however the Adiponectin levels were higher in Dapa alone group. EPC migration improved significantly in both Dapa alone (p=0.05) and Combo group (p=0.05) vs Placebo. Interestingly, urine exosome-based podocyte specific protein such as podocalyxin was actually higher in Combo vs Control. Conclusions: Several parameters showed significant improvement with both Dapa and Combo vs placebo. However, other than glycemic control the Combo didn’t offer any further benefit over Dapa alone in terms of HSC, biochemistry and podocyte markers. Of note, the combination may have deleterious effect on renal podocyte inflammation which needs to be examined in a larger study.

Abstract 3474: Identification of the selective CBL-b inhibitors which effectively prevent T cells from exhaustion and demonstrate synergistic anti-tumor activity in combination with an anti-PD1 antibody

Abstract The E3 ubiquitin ligase Casitas B-lineage lymphoma b (Cbl-b) is a member of the highly conserved family of Cbl (casitas b-lineage lymphoma) proteins. CBL-b is widely expressed in immune cells and plays as a “gate keeper” to negatively regulate the activation of immune cells. Engagement of TCR/BCR triggers the Cbl-b-mediated ubiquitination and degradation of the key components in TCR axis, such as Vav1 and PLCγ, and consequently attenuates TCR/BCR signal axis which finally results in T/B cells exhaustion. Besides, Cbl-b was reported to negatively regulate the activation of dendritic cells, NK, NKT, and mast cells. Cbl-b KO mice show resistance to tumor transplantation, and the anti-tumor activities depend on the T and NK cells. According to the above observation, Cbl-b emerges as a novel intracellular immune checkpoint target to prevent immune cells from exhaustion and improve the antitumor immunotherapy. Here we identified a series of highly potent Cbl-b inhibitors. The inhibition of Cbl-b protein was evaluated by an in vitro CBL-b/E2-ubiquitin assay. Results showed that our candidates strongly inhibited Cbl-b with IC50s at single-digital nanomolar level, and meanwhile, these compounds displayed over than 10-fold selectivity against c-CBL/E2-ubiquitin, a member of the Cbl family, which was reported to synergistically enhance toxicity caused by Cbl-b. Consistent with the Cbl-b inhibition, these compounds further enhanced activation of Jurkat T cells as well as human PBMCs with dramatical increase of IL-2 or IFNɣ secretion in presence of anti-CD3/CD28, and without any significant cytotoxicity even up to 10 µM concertation. Furthermore, these compounds exhibited the restoring function in exhausted T cells in presence with PEG2 or adenosine treatment. Favorable ADME and bioavailability supported the oral administration. In naïve mice, our inhibitors dose-dependently promoted the cytokine IL-2 and IFNɣ production in serum and spleens following the anti-CD3 challenge. In a syngeneic model, CT26, oral administration of our Cbl-b inhibitor effectively inhibited tumor growth, while completely inhibited tumor growth in combination with an anti-PD1 antibody. Further study demonstrated that the increased lymphocytes infiltration and cytokines release in tumor tissues were observed either in Cbl-b mono or in anti-PD1 combo groups, which correlated well with anti-tumor efficacy. In conclusion, our selective Cbl-b inhibitors demonstrated robust immune activation and anti-tumor activity in vitro and in vivo and showed synergistic activities with immune checkpoint inhibitors, such as anti-PD(L)1. Currently, one of our leading molecules is in IND enabling studies. Citation Format: Feng Zhou, Zhengtao Li, Yajing Liu, Guimei Yang, Wansun Mai, Wenqing Yang, Liting Xue, Ping Chen, Renhong Tang. Identification of the selective CBL-b inhibitors which effectively prevent T cells from exhaustion and demonstrate synergistic anti-tumor activity in combination with an anti-PD1 antibody [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3474.

PD04-08 BALIXAFORTIDE, A CXCR4 INHIBITOR, SYNERGIZES WITH DOCETAXEL TO DIMINISH TUMOR GROWTH IN A PROSTATE CANCER MODEL OF BONE METASTASIS

You have accessJournal of UrologyCME1 Apr 2023PD04-08 BALIXAFORTIDE, A CXCR4 INHIBITOR, SYNERGIZES WITH DOCETAXEL TO DIMINISH TUMOR GROWTH IN A PROSTATE CANCER MODEL OF BONE METASTASIS Tyler Robinson, June Escara-Wilke, Jinlu Dai, Ann Arbor, MI, Russell Taichman, Johann Zimmermann, Allschwil Switzerland, and Evan Keller Tyler RobinsonTyler Robinson More articles by this author , June Escara-WilkeJune Escara-Wilke More articles by this author , Jinlu DaiJinlu Dai More articles by this author , Ann ArborAnn Arbor More articles by this author , MI MI More articles by this author , Russell TaichmanRussell Taichman More articles by this author , Johann ZimmermannJohann Zimmermann More articles by this author , Allschwil SwitzerlandAllschwil Switzerland More articles by this author , and Evan KellerEvan Keller More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003228.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Prostate cancer (PCa) bone metastases have been shown to be more resistant to docetaxel than soft tissue metastases. The proinflammatory chemokine receptor CXCR4 has been shown to confer resistance to docetaxel (DOC) in PCa cells. Balixafortide (BLX) is an epitope mimetic inhibitor of CXCR4. Accordingly, we hypothesized and evaluated if BLX would enhance docetaxel-mediated antitumor activity in PCa bone metastases. METHODS: PC-3 luciferase-labeled cells were injected into the tibia of mice to model bone metastases. Four treatment groups were created: vehicle (1% DMSO in DPBS), DOC (5mg/kg), BLX (20mg/kg), and Combo (receiving both DOC and BLX). Mice were injected twice daily subcutaneously with either vehicle or BAL starting on day 1 and weekly intraperitoneally with DOC starting on day one. Tumor burden was measured weekly via bioluminescent imaging. At end of study (29 days), radiographs were taken of the tibiae and blood was collected. Serum levels of TRAcP, IL2, and IFNγ levels were measured using ELISA. Harvested tibiae were decalcified and stained for Ki67, cleaved caspase-3, and CD34 positive cells or microvessels were quantified. Statistical analysis was performed using ANOVA or Student’s t-test. RESULTS: Tumor burden was lower in the Combo group compared to the individual treatment groups. The Combo group had fewer mice with large bone lesions and the lowest TRAcP levels compared to the other groups. No significant difference in Ki67 staining was found among the groups; whereas, cleaved caspase-3 staining was lowest in the Combo group and highest in the BLX. The DOC and Combo groups had more CD34+ microvessels than the control and BLX groups. There was no difference between the treatment groups for IL-2, but the Combo group had increased levels of IFNγ compared to other groups. CONCLUSIONS: Our data demonstrate that that a combination of BLX and DOC has greater anti-tumor activity in a model of PCa bone metastases than either drug alone. The anti-tumor activity was also associated with decreased tumor-induced bone lesions. The mechanism for the anti-tumor activity is not completely elucidated at this time but does not appear to be through impacting proliferation. The observation that IFNγ was increased in the Combo group compared to other groups suggests that immune cell activation contributes to the anti-tumor effect. These data support further evaluation of this combination in metastatic PCa. Source of Funding: This work was supported by Spexis Ltd © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e145 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Tyler Robinson More articles by this author June Escara-Wilke More articles by this author Jinlu Dai More articles by this author Ann Arbor More articles by this author MI More articles by this author Russell Taichman More articles by this author Johann Zimmermann More articles by this author Allschwil Switzerland More articles by this author Evan Keller More articles by this author Expand All Advertisement PDF downloadLoading ...

Curcumin Improves Cytokines and HDL in Diabetic Wistar Rats

In type 2 diabetes mellitus, lipid profile is often found in the form of high-density lipoprotein (HDL) decrease. TNFa and IL-6 are proinflammatory cytokines that are elevated in the blood in many patients with diabetes mellitus. Turmeric is one of the natural ingredients that has anti-cholesterol and anti-inflammatory effects through the work of its active substance, curcumin. This study aimed to evaluate the effect of turmeric extract combined with metformin on HDL, TNFa, and IL-6 levels in diabetic rats. Twenty-four Wistar rats were injected intraperitoneally with streptozotocin and niacinamide. The diabetic rats then were divided into four groups; control group, metformin group, turmeric group, and combination group. The dose of metformin and turmeric which were given, respectively, 45mg/KgBW/day and 200 mg/KgBW/day. While the combo group was given half dose of each. Blood serum was taken to examine the HDL, IL-6, and TNFa levels. The levels of HDL, IL-6, TNFa on diabetic rats that received the combination of metformin and turmeric extract (p0.05) were the best among of all the groups. This means that the combination of metformin and turmeric is better in increasing HDL, reducing IL-6, and TNFa levels better than metformin or turmeric extract alone.

Abstract P6-10-17: Pharmacological Strategies to Target Circadian Clock Genes in TNBC

Abstract Pharmacological Strategies to Target Circadian Clock Genes in TNBC Yuanzhong Pan, Priya Jayachandran, Evanthia T. Roussos Torres, Steve A. Kay Background: Triple-negative breast cancer (TNBC) remains the most aggressive form of breast cancer and more targeted treatment options remain challenging. Our recent analysis of clinical data showed that circadian clock genes (including positive regulators BMAL1 and CLOCK, and negative regulators CRYs, PERs, and REV-ERBs) play important roles in breast cancer, and most prominently in TNBC. We have also shown in other cancer types that cells with a mesenchymal signature and stem-cell like state rely on BMAL1/CLOCK activity to support proliferation. BMAL1 and CLOCK are transcription factors that have remained “undruggable” to date. Alternatively, our lab developed small molecule compounds that target the negative regulators CRY, REV-ERB and CK2. We are therefore prompted to study if the clock genes can serve as a therapeutic target in TNBC using our small molecules that target clock regulators in TNBC. In addition, BMAL1 and CLOCK require ubiquitin E3 ligases and proteasomes for their transcriptional function, thus we hypothesize proteosome inhibitors may provide synergy to improve response to our novel clock-based therapeutics. Methods: We first used shRNA-mediated gene knockdown (KD) to disrupt the core circadian clock regulator BMAL1 and CLOCK in a panel of TNBC cell lines across different molecular subtypes (MDA-MB-231, MDA-MB-157 and MDA-MB-436, MDA-MB-453, HCC70, HCC1143, BT549, Hs578T). After KD, cell proliferation was quantified using CellTiter-Glo. We then tested our small molecule compounds SHP656—that stabilizes CRY, SR29065 and derivatives—that agonizes REV-ERB, and GO289—an inhibitor of CK2 that stabilizes PER. Based on our knowledge of the clock regulators, we tested drug synergy between SHP1705 and proteasome inhibitors MG132 and Carfilzomib across a range of concentrations. RNA-seq of cells treated with either single drug or drug combination were performed to study the mechanism of the synergistic effect. Results: A subset of the tested TNBC cell lines showed significantly impaired proliferation after BMAL1 or CLOCK KD. All cells in the mesenchymal-like molecular subtype responded to BMAL1/CLOCK KD, which is consistent with our previous data in other cancer types. It also confirmed BMAL1 and CLOCK have the potential to serve as therapeutic targets for mesenchymal-like TNBC. We then tested the clock compounds in our TNBC panel. We found SR29065 and GO289 both inhibit cancer cell proliferation at a clinically relevant EC50. Combination of our novel small molecule SHP1705 with the proteosome inhibitor MG132 and carfilzomib, demonstrated significant synergistic effects in vitro. In order to understand the mechanism of the synergistic effect between clock compounds and proteasome inhibitors, we performed RNA-seq on single drug and combo-treated cells. Differential expression analysis revealed that over two-thousand genes are specifically changed in the combo group. GO analysis showed that these genes are enriched in MYC target genes. Because MYC is also, like BMAL1 and CLOCK, a E-box binding transcription factor, this result implies that the mechanism driving synergy may be due to a disruption of E-box-dependent transcription. Conclusions: Here we showed that the core circadian clock regulator BMAL1 and CLOCK are potential therapeutic targets in mesenchymal-like TNBCs. Using REV-ERB agonists and CK2 inhibitors to target BMAL1/CLOCK transcription activity, we can achieve compound EC50s in single-micromolar range. In vivo experiments in murine models of TNBC are underway to determine the efficacy of single agent and combination therapy with proteosome inhibitors. Given the recently established safety of CRY stabilizers there is great potential for translation of these findings to clinical trials in patients with TNBC. Citation Format: Yuanzhong Pan, Priya Jayachandran, Evanthia T. Roussos Torres, Steve A. Kay. Pharmacological Strategies to Target Circadian Clock Genes in TNBC [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P6-10-17.