Cancer stem cells (CSCs) are a distinct population of cells within tumors with capabilities of self-renewal and tumorigenicity. CSCs play a pivotal role in cancer progression, metastasis, and relapse ...and tumor resistance to cytotoxic therapy. Emerging scientific evidence indicates that CSCs adopt several mechanisms, driven by cellular plasticity, senescence and quiescence, to maintain their self-renewal capability and to resist tumor microenvironmental stress and treatments. These pose major hindrances for CSC-targeting anti-cancer therapies: cell plasticity maintains stemness in CSCs and renders tumor cells to acquire stem-like phenotypes, contributing to tumor heterogeneity and CSC generation; cellular senescence induces genetic reprogramming and stemness activation, leading to CSC-mediated tumor progression and metastasis; cell quienscence facilitates CSC to overcome their intrinsic vulnerabilities and therapeutic stress, inducing tumor relapse and therapy resistance. These mechanisms are subjected to spatiotemporal regulation by hypoxia, CSC niche, and extracellular matrix in the tumor microenvironment. Here we integrate the recent advances and current knowledge to elucidate the mechanisms involved in the regulation of plasticity, senescence and quiescence of CSCs and the potential therapeutic implications for the future.
Green infrastructure aims to restore watershed hydrologic function by more closely mimicking pre-development groundwater recharge and evapotranspiration (ET). Bioretention has become a popular ...stormwater control due to its ability to reduce runoff volume through these pathways. Three bioretention cells constructed in low permeability soils in northeast Ohio were monitored for non-winter quantification of inflow, drainage, ET, and exfiltration. The inclusion of an internal water storage (IWS) zone allowed the three cells to reduce runoff by 59%, 42%, and 36% over the monitoring period, in spite of the tight underlying soils. The exfiltration rate and the IWS zone thickness were the primary determinants of volume reduction performance. Post-construction measured drawdown rates were higher than pre-construction soil vertical hydraulic conductivity tests in all cases, due to lateral exfiltration from the IWS zones and ET, which are not typically accounted for in pre-construction soil testing. The minimum rainfall depths required to produce outflow for the three cells were 5.5, 7.4, and 13.8mm. During events with 1-year design rainfall intensities, peak flow reduction varied from 24 to 96%, with the best mitigation during events where peak rainfall rate occurred before the centroid of the rainfall volume, when adequate bowl storage was available to limit overflow.
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•Hydrology of bioretention cells constructed in low permeability soils was evaluated.•Lateral exfiltration was the dominant pathway providing volume reduction.•Bioretention cells were able to completely abstract runoff during small storms.•Peak flow was mitigated during 1-year design rainfall intensities.•Results support the resilience of bioretention systems to poor underlying soils.
Successful treatment of brain tumors such as glioblastoma multiforme (GBM) is limited in large part by the cumulative dose of Radiation Therapy (RT) that can be safely given and the blood-brain ...barrier (BBB), which limits the delivery of systemic anticancer agents into tumor tissue. Consequently, the overall prognosis remains grim. Herein, we report our pilot studies in cell culture experiments and in an animal model of GBM in which RT is complemented by PEGylated-gold nanoparticles (GNPs). GNPs significantly increased cellular DNA damage inflicted by ionizing radiation in human GBM-derived cell lines and resulted in reduced clonogenic survival (with dose-enhancement ratio of ~1.3). Intriguingly, combined GNP and RT also resulted in markedly increased DNA damage to brain blood vessels. Follow-up in vitro experiments confirmed that the combination of GNP and RT resulted in considerably increased DNA damage in brain-derived endothelial cells. Finally, the combination of GNP and RT increased survival of mice with orthotopic GBM tumors. Prior treatment of mice with brain tumors resulted in increased extravasation and in-tumor deposition of GNP, suggesting that RT-induced BBB disruption can be leveraged to improve the tumor-tissue targeting of GNP and thus further optimize the radiosensitization of brain tumors by GNP. These exciting results together suggest that GNP may be usefully integrated into the RT treatment of brain tumors, with potential benefits resulting from increased tumor cell radiosensitization to preferential targeting of tumor-associated vasculature.
Immunologically-cold tumors including glioblastoma (GBM) are refractory to checkpoint blockade therapy, largely due to extensive infiltration of immunosuppressive macrophages (Mϕs). Consistent with a ...pro-tumor role of IL-6 in alternative Mϕs polarization, we here show that targeting IL-6 by genetic ablation or pharmacological inhibition moderately improves T-cell infiltration into GBM and enhances mouse survival; however, IL-6 inhibition does not synergize PD-1 and CTLA-4 checkpoint blockade. Interestingly, anti-IL-6 therapy reduces CD40 expression in GBM-associated Mϕs. We identify a Stat3/HIF-1α-mediated axis, through which IL-6 executes an anti-tumor role to induce CD40 expression in Mϕs. Combination of IL-6 inhibition with CD40 stimulation reverses Mϕ-mediated tumor immunosuppression, sensitizes tumors to checkpoint blockade, and extends animal survival in two syngeneic GBM models, particularly inducing complete regression of GL261 tumors after checkpoint blockade. Thus, antibody cocktail-based immunotherapy that combines checkpoint blockade with dual-targeting of IL-6 and CD40 may offer exciting opportunities for GBM and other solid tumors.
Glioblastomas exhibit vast inter- and intra-tumoral heterogeneity, complicating the development of effective therapeutic strategies. Current in vitro models are limited in preserving the cellular and ...mutational diversity of parental tumors and require a prolonged generation time. Here, we report methods for generating and biobanking patient-derived glioblastoma organoids (GBOs) that recapitulate the histological features, cellular diversity, gene expression, and mutational profiles of their corresponding parental tumors. GBOs can be generated quickly with high reliability and exhibit rapid, aggressive infiltration when transplanted into adult rodent brains. We further demonstrate the utility of GBOs to test personalized therapies by correlating GBO mutational profiles with responses to specific drugs and by modeling chimeric antigen receptor T cell immunotherapy. Our studies show that GBOs maintain many key features of glioblastomas and can be rapidly deployed to investigate patient-specific treatment strategies. Additionally, our live biobank establishes a rich resource for basic and translational glioblastoma research.
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•Rapid establishment of glioblastoma organoids (GBOs) in a defined medium with biobank•GBOs maintain parental tumor cellular heterogeneity, gene expression, and mutations•GBO transplantation exhibits efficient engraftment and aggressive infiltration•Tumor-specific treatment responses in GBOs to drugs and CAR-T cells
A rapid and reliable method to generate patient-derived glioblastoma organoids captures the features and diversity of their respective parental tumors to allow for testing personalized therapies correlated to organoid profile and the establishment of a biobank for further basic and translational glioblastoma research.
: Circulating tumor cells (CTC) are known to be present in the blood of patients with glioblastoma (GBM). Here we report that GBM-derived CTC possess a cancer stem cell (CSC)-like phenotype and ...contribute to local tumorigenesis and recurrence by the process of self-seeding. Genetic probes showed that mouse GBM-derived CTC exhibited Sox2/ETn transcriptional activation and expressed glioma CSC markers, consistent with robust expression of stemness-associated genes including SOX2, OCT4, and NANOG in human GBM patient-derived samples containing CTC. A transgenic mouse model demonstrated that CTC returned to the primary tumor and generated new tumors with enhanced tumorigenic capacity. These CTCs were resistant to radiotherapy and chemotherapy and to circulation stress-induced cell apoptosis. Single-cell RNA-seq analysis revealed that Wnt activation induced stemness and chemoresistance in CTC. Collectively, these findings identify GBM-derived CTC as CSC-like cells and suggest that targeting Wnt may offer therapeutic opportunities for eliminating these treatment-refractory cells in GBM. SIGNIFICANCE: These findings identify CTCs as an alternative source for
tumor invasion and recurrence through local micrometastasis, warranting eradication of systemic "out-of-tumor" CTCs as a promising new therapeutic opportunity for GBM.
Urban stormwater is a substantial source of non-point source pollution. Despite considerable monitoring efforts, little is known about stormwater quality in certain geographic regions. These spatial ...gaps induce uncertainty when extrapolating data and reduce model calibration capabilities, thereby limiting pollutant load reduction strategies. In this study, stormwater quality was monitored from 15 watersheds to characterize pollutant event mean concentrations (EMCs) and loads as a function of urban and forested (i.e., surrogates for pre-development) land use and land covers (LULCs) and rainfall patterns from a geographic region where these data are sparse. Residential and heavy industrial, heavy industrial, and industrial and commercial LULCs, respectively, were the primary generators of nutrients, total suspended solids (TSS), and heavy metals. Increased rainfall intensities (average and peak) significantly increased the EMCs of all particulate bound pollutants. Pollutant loads increased with rainfall depth and, in general, did not follow the same LULC trends as EMCs, suggesting loads were influenced substantially by watershed hydrologic responses. Mean annual urban loads of total phosphorus, total nitrogen, TSS, and zinc (Zn) ranged from 0.4 (low density residential LDR) to 1.5 (heavy industrial), 3.2 (single family residential SFR) to 11.5 (heavy industrial), 122.6 (SFR) to 1219.9 (heavy industrial), and 0.1 (LDR) to 0.7 (commercial) kg/ha/yr, respectively. Annual urban loads of TSS were 3.5 to 34 and − 1.5 to 6.8-fold greater than annual loads from forested and agricultural watersheds, respectively. Mean annual loads of heavy metals from urban LULCs were substantially greater than loads produced by forested and agricultural watersheds (e.g., 8.6 to 92 and 6.8 to 73-fold greater, respectively, for Zn), while loads of nutrients were generally similar between urban and agricultural watersheds. Findings herein suggest non-point source pollution will continue to threaten surface water quality as land is developed; results can help guide the development of cost-efficient stormwater management strategies.
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•Regional data gaps problematic for stormwater quality modeling•Flow proportional runoff samples collected from single land use watersheds in Ohio.•Land use, rainfall intensity were significant influencers of stormwater quality.•Annual urban TSS loads 3.5–34 and -1.5-6.8-fold greater than forested, agricultural.•Surface waters threatened as forested, agricultural land converted to urban.
Blood tests to detect circulating tumor cells (CTC) offer great potential to monitor disease status, gauge prognosis, and guide treatment decisions for patients with cancer. For patients with brain ...tumors, such as aggressive glioblastoma multiforme, CTC assays are needed that do not rely on expression of cancer cell surface biomarkers like epithelial cell adhesion molecules that brain tumors tend to lack. Here, we describe a strategy to detect CTC based on telomerase activity, which is elevated in nearly all tumor cells but not normal cells. This strategy uses an adenoviral detection system that is shown to successfully detect CTC in patients with brain tumors. Clinical data suggest that this assay might assist interpretation of treatment response in patients receiving radiotherapy, for example, to differentiate pseudoprogression from true tumor progression. These results support further development of this assay as a generalized method to detect CTC in patients with cancer.
Immunotherapies have become the standard treatment for melanoma. To further improve patient responses, combinations of immunotherapies and radiotherapy (RT) are being studied, since radiotherapies ...can potentially provide additional immune stimulation, in addition to direct antitumor effects. FLASH-RT is a novel, ultrahigh dose rate, radiation delivery approach, with the potential of at least equivalent tumor control efficacy and reduced damage to healthy tissue. However, the effects of combining FLASH-RT and immunotherapy have not been extensively studied in melanoma. Toll-like receptor (TLR) agonists, such as imiquimod (IMQ), are potent immunostimulatory agents, although their utility is limited due to poor solubility and systemic side effects. We therefore developed a novel combination therapy for melanoma consisting of IMQ delivered to the tumor via a radiopaque and radiation responsive hydrogel combined with FLASH-RT. We found that FLASH was able to effectively stimulate IMQ release from the hydrogel. In addition, we found that the combination of FLASH and released IMQ resulted in synergistic melanoma cell killing in vitro. The combination therapy reduced tumor growth compared to controls, enhanced survival, and resulted in remarkable enhancements in certain tumor cytokine levels. CT imaging allowed the hydrogel to be monitored in vivo. In addition, no adverse effects of the treatment were observed. Overall, this IMQ-gel and FLASH-RT combination may have potential as an improved treatment for melanoma and indicates that the interactions of FLASH-RT and TLR agonists merit further study.Immunotherapies have become the standard treatment for melanoma. To further improve patient responses, combinations of immunotherapies and radiotherapy (RT) are being studied, since radiotherapies can potentially provide additional immune stimulation, in addition to direct antitumor effects. FLASH-RT is a novel, ultrahigh dose rate, radiation delivery approach, with the potential of at least equivalent tumor control efficacy and reduced damage to healthy tissue. However, the effects of combining FLASH-RT and immunotherapy have not been extensively studied in melanoma. Toll-like receptor (TLR) agonists, such as imiquimod (IMQ), are potent immunostimulatory agents, although their utility is limited due to poor solubility and systemic side effects. We therefore developed a novel combination therapy for melanoma consisting of IMQ delivered to the tumor via a radiopaque and radiation responsive hydrogel combined with FLASH-RT. We found that FLASH was able to effectively stimulate IMQ release from the hydrogel. In addition, we found that the combination of FLASH and released IMQ resulted in synergistic melanoma cell killing in vitro. The combination therapy reduced tumor growth compared to controls, enhanced survival, and resulted in remarkable enhancements in certain tumor cytokine levels. CT imaging allowed the hydrogel to be monitored in vivo. In addition, no adverse effects of the treatment were observed. Overall, this IMQ-gel and FLASH-RT combination may have potential as an improved treatment for melanoma and indicates that the interactions of FLASH-RT and TLR agonists merit further study.
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