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Optical-based nanothermometry represents a transformative approach for precise temperature measurements at the nanoscale, which finds versatile applications across biology, medicine, and electronics. The assembly of ratiometric fluorescent 40 nm nanoparticles... 

Nanoparticle-based imaging agents have gained massive attention for the targeted imaging of early-stage cancer. Among these, organic dye-entrapped/assembled nanoparticles have been recognized as potential imaging agents. However, they are limited by poor brightness, low stability, low reproducibil-ity and scalability, and selective surface engineering, which limits their translational potential. The mo-lecular assembly of amphiphilic precursor molecules and the chosen fluorophore can augment the brightness and stability of engineered nanoimaging agents. Herein, we describe an original engineering method for cancer cell membrane-covered ICG-cellulose acetate nanospheres (180 nm) as biomimetic ultra-bright nanoimaging agents for cancer cell imaging. The targeted cancer cell imaging is compared with folic acid-attached ICG-cellulose acetate nanospheres. Encapsulation of fluorescent organic mole-cules (660 dye molecules/ per nanoparticle) in the core of a polymeric network enhances the overall brightness and long-term photostability due to the entrapment of the loaded fluorescent cargo and poor permeation of oxygen to oxidize the dye. The amphiphilic nature of the selected polymeric network accommodates both hydrophilic and hydrophobic cargo molecules (e.g., imaging and therapeutics). The engineered fluorescent nanoparticles exhibited high brightness (780-980 MESF), uniform particle size distribution (180-240 nm), high stability (tested up to 90 days), good biocompatibility with normal cells (95 %), and high scalability (600 mL/batch). For targeted chemotherapeutics, DOX-loaded bio-mimetic nanoparticles demonstrate better chemotherapeutic response (more than 95 % cancer cell death) than folic acid-attached DOX-loaded nanoparticles (78 % cancer cell death) as identified with 24 h MTT assay. The engineered nanoparticles exhibited cancer cell imaging and therapeutics capabili-ties by delivering imaging and drug molecules in cancer mimicked environment in vitro. Our findings suggest that the engineered nanoparticles not only overcome the limitations of nano-imaging but also provide additional advantages for targeted cancer therapeutics. 

Harnessing the power of mesoporous silica to encapsulate organic fluorescent dyes has led to the creation of an extraordinary class of nanocomposite photonic materials. These materials stand out for their ability to produce the brightest fluorescent particles known today, surpassing even the luminosity of quantum dots of similar spectrum and size. The synthesis of these materials offers precise control over the shape and size of the particles, ranging from the nano to the multi-micron scale. Just physical encapsulation of the dyes opens new possibilities for mixing different dyes within individual particles, paving the way for nearly limitless multiplexing capabilities. Moreover, this approach lays the groundwork for the development of highly sensitive sensors capable of detecting subtle changes in temperature and acidity at the nanoscale, among other parameters. This mini-review highlights the mechanism of synthesis, explains the nature of ultrabrightness, and describes the recent advancements and future prospects in the field of ultrabright fluorescent mesoporous silica particles, showcasing their potential for various applications. 

Solid tumors, especially those with aberrant MYCN activation, often harbor an immunosuppressive microenvironment to fuel malignant growth and trigger treatment resistance. Despite this knowledge, there are no effective strategies to tackle this problem. We found that chemokine-like factor (CKLF) is highly expressed by various solid tumor cells and transcriptionally up-regulated by MYCN. Using the MYCN-driven high-risk neuroblastoma as a model system, we demonstrated that as early as the premalignant stage, tumor cells secrete CKLF to attract CCR4-expressing CD4⁺cells, inducing immunosuppression and tumor aggression. Genetic depletion of CD4⁺T regulatory cells abolishes the immunorestrictive and protumorigenic effects of CKLF. Our work supports that disrupting CKLF-mediated cross-talk between tumor and CD4⁺suppressor cells represents a promising immunotherapeutic approach to battling MYCN-driven tumors. 

Here we address an important roadblock that prevents the use of bright fluorescent nanoparticles as individual ratiometric sensors: the possible variation of fluorescence spectra between individual nanoparticles. Ratiometric measurements using florescent dyes have shown their utility in measuring the spatial distribution of temperature, acidity, and concentration of various ions. However, the dyes have a serious limitation in their use as sensors; namely, their fluorescent spectra can change due to interactions with the surrounding dye. Encapsulation of the d, e in a porous material can solve this issue. Recently, we demonstrated the use of ultrabright nanoporous silica nanoparticles (UNSNP) to measure temperature and acidity. The particles have at least two kinds of encapsulated dyes. Ultrahigh brightness of the particles allows measuring of the signal of interest at the single particle level. However, it raises the problem of spectral variation between particles, which is impossible to control at the nanoscale. Here, we study spectral variations between the UNSNP which have two different encapsulated dyes: rhodamine R6G and RB. The dyes can be used to measure temperature. We synthesized these particles using three different ratios of the dyes. We measured the spectra of individual nanoparticles and compared them with simulations. We observed a rather small variation of fluorescence spectra between individual UNSNP, and the spectra were in very good agreement with the results of our simulations. Thus, one can conclude that individual UNSNP can be used as effective ratiometric sensors. 

Hyperactive sphingosine 1-phosphate (S1P) signaling is associated with a poor prognosis of triple-negative breast cancer (TNBC). Despite recent evidence that links the S1P receptor 1 (S1P1) to TNBC cell survival, its role in TNBC invasion and the underlying mechanisms remain elusive. Combining analyses of human TNBC cells with zebrafish xenografts, we found that phosphorylation of S1P receptor 1 (S1P1) at threonine 236 (T236) is critical for TNBC dissemination. Compared to luminal breast cancer cells, TNBC cells exhibit a significant increase of phospho-S1P1 T236 but not the total S1P1 levels. Misexpression of phosphorylation-defective S1P1 T236A (alanine) decreases TNBC cell migration in vitro and disease invasion in zebrafish xenografts. Pharmacologic disruption of S1P1 T236 phosphorylation, using either a pan-AKT inhibitor (MK2206) or an S1P1 functional antagonist (FTY720, an FDA-approved drug for treating multiple sclerosis), suppresses TNBC cell migration in vitro and tumor invasion in vivo. Finally, we show that human TNBC cells with AKT activation and elevated phospho-S1P1 T236 are sensitive to FTY720-induced cytotoxic effects. These findings indicate that the AKT-enhanced phosphorylation of S1P1 T236 mediates much of the TNBC invasiveness, providing a potential biomarker to select TNBC patients for the clinical application of FTY720. 

Despite the development of metabolism-based therapy for a variety of malignancies, resistance to single-agent treatment is common due to the metabolic plasticity of cancer cells. Improved understanding of how malignant cells rewire metabolic pathways can guide the rational selection of combination therapy to circumvent drug resistance. Here, we show that human T-ALL cells shift their metabolism from oxidative decarboxylation to reductive carboxylation when the TCA cycle is disrupted. The α-ketoglutarate dehydrogenase complex (KGDHC) in the TCA cycle regulates oxidative decarboxylation by converting α-ketoglutarate (α-KG) to succinyl-CoA, while isocitrate dehydrogenase (IDH) 1 and 2 govern reductive carboxylation. Metabolomics flux analysis of T-ALL reveals enhanced reductive carboxylation upon genetic depletion of the E2 subunit of KGDHC, dihydrolipoamide-succinyl transferase (DLST), mimicking pharmacological inhibition of the complex. Mechanistically, KGDHC dysfunction causes increased demethylation of nuclear DNA by α-KG-dependent dioxygenases (e.g., TET demethylases), leading to increased production of both IDH1 and 2. Consequently, dual pharmacologic inhibition of the TCA cycle and TET demethylases demonstrates additive efficacy in reducing the tumor burden in zebrafish xenografts. These findings provide mechanistic insights into how T-ALL develops resistance to drugs targeting the TCA cycle and therapeutic strategies to overcome this resistance. 

Screening for drugs that disrupt embryonic development in zebrafish can help identify treatments that suppress metastasis. 

Abstract Triple-negative breast cancer (TNBC) is traditionally considered a glycolytic tumor with a poor prognosis while lacking targeted therapies. Here we show that high expression of dihydrolipoamide S-succinyltransferase (DLST), a tricarboxylic acid (TCA) cycle enzyme, predicts poor overall and recurrence-free survival among TNBC patients. DLST depletion suppresses growth and induces death in subsets of human TNBC cell lines, which are capable of utilizing glutamine anaplerosis. Metabolomics profiling reveals significant changes in the TCA cycle and reactive oxygen species (ROS) related pathways for sensitive but not resistant TNBC cells. Consequently, DLST depletion in sensitive TNBC cells increases ROS levels while N-acetyl-L-cysteine partially rescues cell growth. Importantly, suppression of the TCA cycle through DLST depletion or CPI-613, a drug currently in clinical trials for treating other cancers, decreases the burden and invasion of these TNBC. Together, our data demonstrate differential TCA-cycle usage in TNBC and provide therapeutic implications for the DLST-dependent subsets.