Unveiling cancer dormancy: Intrinsic mechanisms and extrinsic forces.

Tumor cells disseminate in various distant organs at early stages of cancer progression. These disseminated tumor cells (DTCs) can stay dormant/quiescent without causing patient symptoms for years or decades. These dormant tumor cells survive despite curative treatments by entering growth arrest, escaping immune surveillance, and/or developing drug resistance. However, these dormant cells can reactivate to proliferate, causing metastatic progression and/or relapse, posing a threat to patients' survival. It's unclear how cancer cells maintain dormancy and what triggers their reactivation. What are better approaches to prevent metastatic progression and relapse through harnessing cancer dormancy? To answer these remaining questions, we reviewed the studies of tumor dormancy and reactivation in various types of cancer using different model systems, including the brief history of dormancy studies, the intrinsic characteristics of dormant cells, and the external cues at the cellular and molecular levels. Furthermore, we discussed future directions in the field and the strategies for manipulating dormancy to prevent metastatic progression and recurrence.

Flavonol profiles of mature leaves allow discriminating Toona sinensis Roem from different north-south geographical origins across China with varied antioxidant activities.

Toona sinensis (A. Juss.) Roem, a multipurpose economic tree, is widely cultivated across Asia, but its high-yielding mature leaves are largely overlooked. This study systematically analysed the flavonols in the mature leaves of T. sinensis from 44 different geographic locations across China, using HPLC-DAD and HPLC-ESI-MS2 techniques. In total, 18 flavonols were detected, among which 6 (f1, f3, f7, f14, f15, and f17) were firstly identified in this plant. Significant variations in quality among different T. sinensis varieties were observed (p < 0.01). Through OPLS-DA analysis, all samples could be clearly categorised into two distinct geographical groups. The northern varieties (N1-N20) exhibited concise flavonol fingerprints with higher total flavonol content (TFC) (727.55 ± 22.79 mg/100 g fresh weight, FW), predominantly non-acylated flavonols (705.95 ± 21.65 mg/100 g FW), particularly quercetin glycosides (614.60 ± 22.76 mg/100 g FW). In contrast, the southern varieties (S1-S24) presented more intricate flavonol profiles with lower TFC (622.81 ± 21.82 mg/100 g FW) and balanced amounts of quercetin (344.75 ± 16.41 mg/100 g FW) and kaempferol glycosides (278.06 ± 12.29 mg/100 g FW). Notably, the southern samples possessed higher content of acylated flavonols (184.50 ± 12.87 mg/100 g FW), especially galloylated ones, which contributed to their heightened antioxidant activities. Quercetin 3-O-rhamnoside (f11') and kaempferol 3-O-galloyglucoside (f11) were determined to be the crucial biomarkers for quality discrimination. Considering quality control of mature T. sinensis leaves as potential resources for natural flavonol extraction, this study suggested that their northern/southern geographic origins should be distinguished first. Additionally, the flavonol profiles allow for discriminating the origin and assessing the quality of T. sinensis.

Tumor removal limits prostate cancer cell dissemination in bone and osteoblasts induce cancer cell dormancy through focal adhesion kinase.

BACKGROUND: Disseminated tumor cells (DTCs) can enter a dormant state and cause no symptoms in cancer patients. On the other hand, the dormant DTCs can reactivate and cause metastases progression and lethal relapses. In prostate cancer (PCa), relapse can happen after curative treatments such as primary tumor removal. The impact of surgical removal on PCa dissemination and dormancy remains elusive. Furthermore, as dormant DTCs are asymptomatic, dormancy-induction can be an operational cure for preventing metastases and relapse of PCa patients. METHODS: We used a PCa subcutaneous xenograft model and species-specific PCR to survey the DTCs in various organs at different time points of tumor growth and in response to tumor removal. We developed in vitro 2D and 3D co-culture models to recapitulate the dormant DTCs in the bone microenvironment. Proliferation assays, fluorescent cell cycle reporter, qRT-PCR, and Western Blot were used to characterize the dormancy phenotype. We performed RNA sequencing to determine the dormancy signature of PCa. A drug repurposing algorithm was applied to predict dormancy-inducing drugs and a top candidate was validated for the efficacy and the mechanism of dormancy induction. RESULTS: We found DTCs in almost all mouse organs examined, including bones, at week 2 post-tumor cell injections. Surgical removal of the primary tumor reduced the overall DTC abundance, but the DTCs were enriched only in the bones. We found that osteoblasts, but not other cells of the bones, induced PCa cell dormancy. RNA-Seq revealed the suppression of mitochondrial-related biological processes in osteoblast-induced dormant PCa cells. Importantly, the mitochondrial-related biological processes were found up-regulated in both circulating tumor cells and bone metastases from PCa patients' data. We predicted and validated the dormancy-mimicking effect of PF-562,271 (PF-271), an inhibitor of focal adhesion kinase (FAK) in vitro. Decreased FAK phosphorylation and increased nuclear translocation were found in both co-cultured and PF-271-treated C4-2B cells, suggesting that FAK plays a key role in osteoblast-induced PCa dormancy. CONCLUSIONS: Our study provides the first insights into how primary tumor removal enriches PCa cell dissemination in the bones, defines a unique osteoblast-induced PCa dormancy signature, and identifies FAK as a PCa cell dormancy gatekeeper.

Characterization of key aroma-active compounds in blue honeysuckle (Lonicera caerulea L.) berries by sensory-directed analysis.

Blue honeysuckle (Lonicera caerulea L.) berries are nutritionally rich and unique in flavor. However, its aroma compounds have not been known well. In this study, the key aroma-active compounds in 8 different varieties of blue honeysuckle berries were studied by sensory-directed analysis. Sensory evaluation suggested that the aroma profile of blue honeysuckle berry was fruity, floral, grassy, sweet, and sour. A total of 68 aroma compounds were detected by two-dimensional comprehensive gas chromatography-olfactometry-mass spectrometry analysis (GC × GC-O-MS). Then, aroma extraction dilution analysis (AEDA) and odor activity value (OAV) showed that 12 compounds were indicated to be the major aroma contributors. According to the principal component analysis (PCA) results, eight varieties were divided into three categories for their differences on alcohols and terpenoids content. Finally, the aroma recombination and omission experiments determined that linalool, hexanal, eucalyptol, octanal, nonanal, and ethyl 2-methylbutyrate were the key aroma-active compounds in blue honeysuckle berries.

Parathyroid Hormone-Related Protein/Parathyroid Hormone Receptor 1 Signaling in Cancer and Metastasis.

PTHrP exerts its effects by binding to its receptor, PTH1R, a G protein-coupled receptor (GPCR), activating the downstream cAMP signaling pathway. As an autocrine, paracrine, or intracrine factor, PTHrP has been found to stimulate cancer cell proliferation, inhibit apoptosis, and promote tumor-induced osteolysis of bone. Despite these findings, attempts to develop PTHrP and PTH1R as drug targets have not produced successful results in the clinic. Nevertheless, the efficacy of blocking PTHrP and PTH1R has been shown in various types of cancer, suggesting its potential for therapeutic applications. In light of these conflicting data, we conducted a comprehensive review of the studies of PTHrP/PTH1R in cancer progression and metastasis and highlighted the strengths and limitations of targeting PTHrP or PTH1R in cancer therapy. This review also offers our perspectives for future research in this field.

Enzalutamide-induced and PTH1R-mediated TGFBR2 degradation in osteoblasts confers resistance in prostate cancer bone metastases.

Enzalutamide resistance has been observed in approximately 50% of patients with prostate cancer (PCa) bone metastases. Therefore, there is an urgent need to investigate the mechanisms and develop strategies to overcome resistance. We observed enzalutamide resistance in bone lesion development induced by PCa cells in mouse models. We found that the bone microenvironment was indispensable for enzalutamide resistance because enzalutamide significantly inhibited the growth of subcutaneous C4-2B tumors and the proliferation of C4-2B cells isolated from the bone lesions, and the resistance was recapitulated only when C4-2B cells were co-cultured with osteoblasts. In revealing how osteoblasts contribute to enzalutamide resistance, we found that enzalutamide decreased TGFBR2 protein expression in osteoblasts, which was supported by clinical data. This decrease was possibly through PTH1R-mediated endocytosis. We showed that PTH1R blockade rescued enzalutamide-mediated decrease in TGFBR2 levels and enzalutamide responses in C4-2B cells that were co-cultured with osteoblasts. This is the first study to reveal the contribution of the bone microenvironment to enzalutamide resistance and identify PTH1R as a feasible target to overcome the resistance in PCa bone metastases.

Development of a system dynamics model for dioxin flow and its application to the energy sector.

Dioxin pollution has been problematic in Taiwan. Although the government has established emission standards and emission inventory to control dioxin pollution, such efforts only apply to pollution emissions; no attempt has been made to understand the flow of dioxins in different environmental media. In this study, the STELLA software was used to model the flow pattern of dioxins in various media. This model and the RAIDAR model established by the Canadian Environmental Model Research Center were used to simulate dioxin flow in Taiwan, and their results were compared with the measured values. The accuracies of the RAIDAR and STELLA models were 63.92% and 49.78%, respectively. This shows that the simulation with the STELLA model provided results closer to the measured values and that the error was less than ten times that of the RAIDAR model, indicating that the proposed model is predictive. In addition, we used the results of a system dynamics model for dioxin flow and an air resource co-benefits (ARCoB) model to apply the obtained results to the energy sector to quantify the co-benefits of reducing dioxin, greenhouse gas, and air pollutant emissions on the basis of the policy target for the year 2030. The total co-benefits of natural gas and renewable energy (RE) scenarios were US$9.63 billion and US$12.57 billion, respectively; the benefit-cost ratios were 2.89 and 20.67, respectively. The development of an RE policy as an alternative to a coal-fired power generation policy will contribute to the best co-benefits of integrated reductions and will also contribute to human health.

Dive into Single, Seek Out Multiple: Probing Cancer Metastases via Single-Cell Sequencing and Imaging Techniques.

Metastasis is the cause of most cancer deaths and continues to be the biggest challenge in clinical practice and laboratory investigation. The challenge is largely due to the intrinsic heterogeneity of primary and metastatic tumor populations and the complex interactions among cancer cells and cells in the tumor microenvironment. Therefore, it is important to determine the genotype and phenotype of individual cells so that the metastasis-driving events can be precisely identified, understood, and targeted in future therapies. Single-cell sequencing techniques have allowed the direct comparison of the genomic and transcriptomic changes among different stages of metastatic samples. Single-cell imaging approaches have enabled the live visualization of the heterogeneous behaviors of malignant and non-malignant cells in the tumor microenvironment. By applying these technologies, we are achieving a spatiotemporal precision understanding of cancer metastases and clinical therapeutic translations.

Differences and Correlations of Morphological Characteristics and Fatty Acid Profiles of Seeds of Toona sinensis.

Toona sinensis (A.Juss.) M.Roem., a multi-purpose tree of Meliaceae, is widely distributed and intensively cultivated in Asia, yet its high yielding, lipid-rich seeds are rarely exploited. The present study systematically analyzed the differences and correlations of seed morphological characteristics and fatty acid (FA) profiles of 62 representative T. sinensis germplasms distributed across northern to southern China. T. sinensis seeds were rich in total FAs (TFA, 107.03-176.18 mg/g). Additionally, linoleic acid (54.69-100.59 mg/g), α-linolenic acid (ALA, 22.47-45.02 mg/g), oleic acid (OA, 5.12-23.94 mg/g), palmitic acid (6.87-14.14 mg/g), stearic acid (SA, 3.13-6.57 mg/g) and elaidic acid (1.70-2.88 mg/g) were the major FAs measured by GC/MS analysis. Size (average width of 3.94±0.01 mm and length of 5.79±0.02 mm) and mass (average thousand-seed weight of 10.52±0.17 g) were greater in T. sinensis seeds collected south than north of 30° latitude. These traits were also positively correlated with unsaturated FA content and negatively related to SA and saturated FA contents (P<0.05). Significant positive correlations were found between seed length and polyunsaturated FA (R2 =0.370) and ALA levels (R2 =0.296), as well as between thousand-seed weight and monounsaturated FAs (R2 =0.309) and OA levels (R2 =0.297) (P<0.05). Seventeen T. sinensis germplasms gathered by cluster analysis as cluster IV were determined as desirable for oil processing due to their higher TFA and ALA contents and greater seed size and mass than others. Generally, the wider, heavier, and especially longer seeds of T. sinensis contain much higher levels of FAs, especially ALA, and are the more promising sources for breeding and the oil processing industry.

Comparative and Phylogenetic Analyses of the Complete Chloroplast Genomes of Six Almond Species (Prunus spp. L.).

As a source of genetic variation, almond germplasm resources are of great significance in breeding. To better reveal the mutation characteristics and evolution patterns of the almond chloroplast (cp) genome, the complete cp genomes from six almond species were analyzed. The lengths of the chloroplast genome of the six almond species ranged from 157,783 bp to 158,073 bp. For repeat sequence analysis, 53 pairs of repeats (30 bp or longer) were identified. A total of 117 SSR loci were observed, including 96 polymorphic SSR loci. Nine highly variable regions with a nucleotide variability (Pi) higher than 0.08, including rps16, rps16-psbK, atpF-atpH, rpoB, ycf3-rps4, rps4-ndhJ, accD-psaI and rps7-orf42 (two highly variable regions) were located. Based on the chloroplast genome evolution analysis, three species (P. tenella, P. pedunculata and P. triloba) and wild cherry (P. tomentosa) were grouped into clade I. Clade II consisted of two species (P. mongolica and P. tangutica) and wild peach (P. davidiana). Clade III included the common almond (P. dulcis), cultivated peach (P. persica) and GanSu peach (P. kansuensis). This result expands the researchers' vision of almond plant diversity and promotes an understanding of the evolutionary relationship among almond species. In brief, this study provides abundant resources for the study of the almond chloroplast genome, and has an important reference value for study of the evolution and species identification of almond.

Glutathionyl-S-chlorogenic acid is present in fruit of Vaccinium species, potato tubers and apple juice.

A hydroxycinnamate-like component was identified in highbush blueberry (Vaccinium corymbosum) fruit, which had identical UV and mass spectrometric properties to an S-linked glutathionyl conjugate of chlorogenic acid synthesized using a peroxidase-catalyzed reaction. The conjugate was present in fruits from all highbush blueberry genotypes grown in one season, reaching 7-20% of the relative abundance of 5-caffeoylquininc acid. It was enriched, along with anthocyanins, by fractionation on solid phase cation-exchange units. Mining of pre-existing LC-MS data confirmed that this conjugate was ubiquitous in highbush blueberries, but also present in other Vaccinium species. Similar data mining identified this conjugate in potato tubers with enrichment in peel tissues. In addition, the conjugate was also present in commercial apple juice and was stable to pasteurization and storage. Although glutathionyl conjugates of hydroxycinnamic acids have been noted previously, this is the first report of glutathionyl conjugates of chlorogenic acids in commonly-eaten fruits and vegetables.

Distinct physiological and transcriptional responses of leaves of paper mulberry (Broussonetia kazinoki × B. papyrifera) under different nitrogen supply levels.

Paper mulberry, a vigorous pioneer species used for ecological reclamation and a high-protein forage plant for economic development, has been widely planted in China. To further develop its potential value, it is necessary to explore the regulatory mechanism of nitrogen metabolism for rational nitrogen utilization. In this study, we investigated the morphology, physiology and transcriptome of a paper mulberry hybrid (Broussonetia kazinoki × B. papyrifera) in response to different nitrogen concentrations. Moderate nitrogen promoted plant growth and biomass accumulation. Photosynthetic characteristics, concentration of nitrogenous compounds and activities of enzymes were stimulated under nitrogen treatment. However, these enhancements were slightly or severely inhibited under excessive nitrogen supply. Nitrite reductase and glutamate synthase were more sensitive than nitrate reductase and glutamine synthetase and more likely to be inhibited under high nitrogen concentrations. Transcriptome analysis of the leaf transcriptome identified 161,961 unigenes. The differentially expressed genes associated with metabolism of nitrogen, alanine, aspartate, glutamate and glycerophospholipid showed high transcript abundances after nitrogen application, whereas those associated with glycerophospholipid, glycerolipid, amino sugar and nucleotide sugar metabolism were down-regulated. Combined with weighted gene coexpression network analysis, we uncovered 16 modules according to similarity in expression patterns. Asparagine synthetase and inorganic pyrophosphatase were considered two hub genes in two modules, which were associated with nitrogen metabolism and phosphorus metabolism, respectively. The expression characteristics of these genes may explain the regulation of morphological, physiological and other related metabolic strategies harmoniously. This multifaceted study provides valuable insights to further understand the mechanism of nitrogen metabolism and to guide utilization of paper mulberry.

Potent and Preferential Degradation of CDK6 via Proteolysis Targeting Chimera Degraders.

A focused PROTAC library hijacking cancer therapeutic target CDK6 was developed. A design principle as "match/mismatch" was proposed for understanding the degradation profile differences in these PROTACs. Notably, potent PROTACs with specific and remarkable CDK6 degradation potential were generated by linking CDK6 inhibitor palbociclib and E3 ligase CRBN recruiter pomalidomide. The PROTAC strongly inhibited proliferation of hematopoietic cancer cells including multiple myeloma and robustly degraded copy-amplified/mutated forms of CDK6, indicating future potential clinical applications.

Correction to: Developing potent PROTACs tools for selective degradation of HDAC6 protein.

In the original publication the title of X axis in figure 1G is incorrectly published as "Compound (µmol/L)". The correct title of X axis in figure 1G should be read as "Compound (nmol/L)".

Lung Tumorigenesis Alters the Expression of Slit2-exon15 Splicing Variants in Tumor Microenvironment.

Slit2 expression is downregulated in various cancers, including lung cancer. We identified two Slit2 splicing variants at exon15-Slit2-WT and Slit2-ΔE15. In the RT-PCR analyses, the Slit2-WT isoform was predominantly expressed in all the lung cancer specimens and in their normal lung counterparts, whereas Slit2-ΔE15 was equivalently or predominantly expressed in 41% of the pneumothorax specimens. A kRasG12D transgenic mice system was used to study the effects of tumorigenesis on the expressions of the Slit2-exon15 isoforms. The results revealed that a kRasG12D-induced lung tumor increased the Slit2-WT/Slit2-ΔE15 ratio and total Slit2 expression level. However, the lung tumors generated via a tail vein injection of lung cancer cells decreased the Slit2-WT/Slit2-ΔE15 ratio and total Slit2 expression level. Interestingly, the lipopolysaccharide (LPS)-induced lung inflammation also decreased the Slit2-WT/Slit2-ΔE15 ratio. Since Slit2 functions as an anti-inflammatory factor, the expression of Slit2 increases in kRasG12D lungs, which indicates that Slit2 suppresses immunity during tumorigenesis. However, an injection of lung cancer cells via the tail vein and the LPS-induced lung inflammation both decreased the Slit2 expression. The increased Slit2 in the tumor microenvironment was mostly Slit2-WT, which lacks growth inhibitory activity. Thus, the results of our study suggested that the upregulation of Slit2-WT, but not Slit2-ΔE15, in a cancer microenvironment is an important factor in suppressing immunity while not interfering with cancer growth.

Induction of apoptosis in MDA-MB-231 breast cancer cells by a PARP1-targeting PROTAC small molecule.

Poly (ADP-ribose) polymerase-1 (PARP1) is a major member of the PARP superfamily that is involved in DNA damage signalling and other important cellular processes. Here we report the development of a small molecule targeting PARP1 based on the PROTAC strategy. In the MDA-MB-231 cell line, the representative compound 3 can induce significant PARP1 cleavage and programmed cell death.

Selective variable optical attenuator for visible and mid-Infrared wavelengths.

This work demonstrates a variable optical attenuator (VOA) using dynamic scattering mode (DSM) in ion-doped liquid crystals with negative dielectric anisotropy. The mechanism of attenuation comes from optical scattering, which is generated by the electrically induced instability of undulation of LC textures. Electric fields are applied to switch the initial transparent state of the designed VOA to scattering states, varying the transmittance. The electric field also changes the size of the scattering domain from the LC texture and causes the designed device to exhibit an ultra-broadband selective operation in a visible to mid-IR spectral range. Furthermore, the VOA can selectively block one visible or mid-IR wavelength of light while letting other light pass. Such a VOA has many superior optical switching properties, such as high on/off contrast, insensitivity to polarization, and spectral selectivity; therefore, it has the potential to be used in practical optical systems.

Antioxidant capacities and anthocyanin characteristics of the black-red wild berries obtained in Northeast China.

Various edible berries widely accessible in nature in Northeast China are poorly exploited. The compositions and contents of anthocyanins in black (Padus maackii, Padus avium, Lonicera caerulea, and Ribes nigrum) and red (Ribes rubrum, Sambucus williamsii, Rubus idaeus, and Ribes procumbens) wild berries in Northeast China were firstly characterized by HPLC-DAD/ESI-MS(2). Twenty-three anthocyanins were detected and identified. Cyanidin glycosides were dominant in both berries. Six anthocyanins were reported for the first time in P. avium, R. rubrum, and Sambucus. Total anthocyanin content (TAC) ranged from 10mg/100gfreshweight (FW) (R. procumbens) to 1058mg/100gFW (P. maackii) among berries. The TACs and antioxidant activities assessed by DPPH and FRAP assays were much higher in black than in red berries. Black-red berries, especially P. maackii and P. avium, can be used in developing functional foods and in improving breeding programs.

Flavone synthases from Lonicera japonica and L. macranthoides reveal differential flavone accumulation.

Flavones are important secondary metabolites found in many plants. In Lonicera species, flavones contribute both physiological and pharmaceutical properties. However, flavone synthase (FNS), the key enzyme responsible for flavone biosynthesis, has not yet been characterized in Lonicera species. In this study, FNSII genes were identified from Lonicera japonica Thunb. and L. macranthoides Hand.-Mazz. In the presence of NADPH, the recombinant cytochrome P450 proteins encoded by LjFNSII-1.1, LjFNSII-2.1, and LmFNSII-1.1 converted eriodictyol, naringenin, and liquiritigenin to the corresponding flavones directly. The different catalytic properties between LjFNSII-2.1 and LjFNSII-1.1 were caused by a single amino acid substitution at position 242 (glutamic acid to lysine). A methionine at position 206 and a leucine at position 381 contributed considerably to the high catalytic activity of LjFNSII-1.1. In addition, LjFNSII-1.1&2.1 and LmFNSII-1.1 also biosynthesize flavones that were further modified by O-glycosylation in transgenic tobacco. The expression levels of the FNSII genes were consistent with flavone accumulation patterns in flower buds. Our findings suggested that the weak catalytic activity of LmFNSII-1.1 and the relatively low expression of LmFNSII-1.1 in flowers might be responsible for the low levels of flavone accumulation in flower buds of L. macranthoides.