Patients with BRAF-mutant melanoma show substantial responses to combined BRAF and MEK inhibition, but most relapse within 2 years. A major reservoir for drug resistance is minimal residual disease ...(MRD), comprised of drug-tolerant tumor cells laying in a dormant state. Towards exploiting potential therapeutic vulnerabilities of MRD, we established a genetically engineered mouse model of BrafV600E-driven melanoma MRD wherein genetic BrafV600E extinction leads to strong but incomplete tumor regression. Transcriptional time-course analysis after BrafV600E extinction revealed that after an initial surge of immune activation, tumors later became immunologically "cold" after MRD establishment. Computational analysis identified candidate T-cell recruiting chemokines as strongly upregulated initially and steeply decreasing as the immune response faded. Therefore, we hypothesized that sustaining chemokine signaling could impair MRD maintenance through increased recruitment of effector T cells. We found that intratumoral administration of recombinant Cxcl9 (rCxcl9), either naked or loaded in microparticles, significantly impaired MRD relapse in BRAF-inhibited tumors, including several complete pathologic responses after microparticle-delivered rCxcl9 combined with BRAF and MEK inhibition. Our experiments constitute proof of concept that chemokine-based microparticle delivery systems are a potential strategy to forestall tumor relapse and thus improve the clinical success of first-line treatment methods.
Ovarian cancer accounts for 3% of the total cancers in women, yet it is the fifth leading cause of cancer deaths among women. The BRCA1/2 germline and somatic mutations confer a deficiency of the ...homologous recombination (HR) repair pathway. Inhibitors of poly (ADP-ribose) polymerase (PARP), another important component of DNA damage repair, are somewhat effective in BRCA1/2 mutant tumors. However, ovarian cancers often reacquire functional BRCA and develop resistance to PARP inhibitors. Polyamines have been reported to facilitate the DNA damage repair functions of PARP. Given the elevated levels of polyamines in tumors, we hypothesized that treatment with the polyamine synthesis inhibitor, α-difluoromethylornithine (DFMO), may enhance ovarian tumor sensitivity to the PARP inhibitor, rucaparib. In HR-competent ovarian cancer cell lines with varying sensitivities to rucaparib, we show that co-treatment with DFMO increases the sensitivity of ovarian cancer cells to rucaparib. Immunofluorescence assays demonstrated that, in the presence of hydrogen peroxide-induced DNA damage, DFMO strongly inhibits PARylation, increases DNA damage accumulation, and reduces cell viability in both HR-competent and deficient cell lines. In vitro viability assays show that DFMO and rucaparib cotreatment significantly enhances the cytotoxicity of the chemotherapeutic agent, cisplatin. These results suggest that DFMO may be a useful adjunct chemotherapeutic to improve the anti-tumor efficacy of PARP inhibitors in treating ovarian cancer.
BRAF mutations are present in over half of all melanoma tumors. Although BRAF inhibitors significantly improve survival of patients with metastatic melanoma, recurrences occur within several months. ...We previously reported that BRAF mutant melanoma cells are more sensitive to a novel arylmethyl-polyamine (
AP
) compound that exploits their increased polyamine uptake compared to that of BRAF wildtype cells. Using an animal model of BRAF inhibitor-resistant melanoma, we show that co-treatment with the BRAF inhibitor, PLX4720, and
AP
significantly delays the recurrence of PLX4720-resistant melanoma tumors and decreases tumor-promoting macrophages. Development of BRAF inhibitor-resistance enriches for metastatic cancer stem cells (CSC) and increases tumor-promoting macrophages. In vitro studies demonstrated that CD304
+
, CXCR4
+
spheroid cultures of BRAF mutant melanoma cells are resistant to PLX4720 but are more sensitive to
AP
compared to monolayer cultures of the same cells.
AP
significantly inhibited YUMM1.7 melanoma cell invasiveness across a Matrigel-coated filter using the CXCR4 ligand, SDF-1α, as the chemoattractant.
AP
also blocked the chemotactic effect of SDF-1α on CXCR4
+
macrophages and inhibited M2 polarization of macrophages. In melanoma-macrophage co-cultures,
AP
prevented the PLX4720-induced release of pro-tumorigenic growth factors, such as VEGF, from macrophages and prevented the macrophage rescue of BRAF mutant melanoma cells treated with PLX4720. Our study offers a novel therapy (
AP
) to treat chemo-resistant melanoma.
AP
is unique because it targets the polyamine transport system in BRAF inhibitor-resistant CSCs and also blocks CXCR4 signaling in invasive melanoma cells and pro-tumorigenic macrophages.
Abstract
Patients with BRAF-mutant melanoma show significant responses to combined BRAF and MEK inhibition, but most patients relapse within 2 years. A major reservoir for such drug resistance is ...minimal residual disease (MRD), which is comprised of drug-tolerant tumor cells laying in a dormant state. Towards exploiting potential therapeutic vulnerabilities of MRD, we established a genetically engineered mouse model of BrafV600E-driven melanoma MRD wherein genetic BrafV600E extinction leads to strong but incomplete tumor regression. Transcriptional time-course analysis of tumors after BrafV600E extinction revealed that after an initial surge of immune activation, tumors later became immunologically "cold" after MRD establishment. Computational analysis identified candidate T-cell recruiting chemokines that may be central players in the process, being strongly upregulated initially and then steeply decreasing as the immune response faded. As a result, we hypothesized that sustaining the chemokine signaling could impair MRD maintenance through increased recruitment of effector T-cells. We show that intratumoral administration of recombinant Cxcl9, either naked or loaded in microparticles, significantly impaired the relapse of MRD in BRAF-inhibited tumors, including several complete pathological responses after the triple combination of microparticle-delivered rCxcl9 with BRAF and MEK-inhibition. Our experiments constitute a proof of concept that chemokine-based microparticle delivery systems are a potential strategy to forestall tumor relapse and thus improve the clinical success of frontline treatment methods.
Citation Format: Gabriele Romano, Francesca Paradiso, Peng Li, Pooja Shukla3, Lindsay Barger, Olivia El Naggar, John P. Miller, Roger J. Liang, Timothy L. Helms, Jennifer A. Wargo, Francesca Taraballi, James C. Costello, Lawrence N. Kwong. Microparticle-delivered Cxcl9 prolongs Braf inhibitor efficacy in melanoma 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 1853.
Performance of slabs on grade is governed mainly by the mutual coupled performance of the slab stiffness and the subgrade support. The use of reinforcement geogrids to increase the stiffness and ...structural capacity of the subgrade support can be potentially advantageous, but is currently poorly understood, especially under frequent transient loading conditions. The objective of this experimental study is to investigate the beneficial use of geogrids as a reinforcing material in a loose subgrade. This paper presents the results of large scale laboratory tests on slab strips (300 mm wide × 150 mm thick × 2,500 mm long) supported on a reinforced subgrade. The slab strips contained either steel fibers or were reinforced with welded wire fabric (WWF), and are compared with earlier tests on an unreinforced subgrade. All slab strips were tested under a central point load and were restrained from uplift at the ends. Both monotonic and cyclic loads are considered. The geogrid increases the bottom surface cracking load of the slab strip, does not affect the top surface cracking load, and increases the ultimate load carrying capacity. The use of steel fiber reinforced concrete (SFRC) was successful in providing post-cracking strength to maintain slab integrity when compared to the WWF reinforced slab strip used in this investigation. A simplified analysis of beams on grade is carried out for the elastic and post-cracking regions, and predicted values for the SFRC slab strip under monotonic load are compared to the measured load-deformation response. The comparison indicates that the theoretical response underestimates the capacity of the SFRC slab strip.
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