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1.
Chromosoma ; 133(1): 77-92, 2024 01.
Artigo em Inglês | MEDLINE | ID: mdl-37256347

RESUMO

Chromosome gains or losses often lead to copy number variations (CNV) and loss of heterozygosity (LOH). Both quantities are low in hematologic "liquid" cancers versus solid tumors in data of The Cancer Genome Atlas (TCGA) that also shows the fraction of a genome affected by LOH is ~ one-half of that with CNV. Suspension cultures of p53-null THP-1 leukemia-derived cells conform to these trends, despite novel evidence here of genetic heterogeneity and transiently elevated CNV after perturbation. Single-cell DNAseq indeed reveals at least 8 distinct THP-1 aneuploid clones with further intra-clonal variation, suggesting ongoing genetic evolution. Importantly, acute inhibition of the mitotic spindle assembly checkpoint (SAC) produces CNV levels that are typical of high-CNV solid tumors, with subsequent cell death and down-selection to novel CNV. Pan-cancer analyses show p53 inactivation associates with aneuploidy, but leukemias exhibit a weaker trend even though p53 inactivation correlates with poor survival. Overexpression of p53 in THP-1 does not rescue established aneuploidy or LOH but slightly increases cell death under oxidative or confinement stress, and triggers p21, a key p53 target, but without affecting net growth. Our results suggest that factors other than p53 exert stronger pressures against aneuploidy in liquid cancers, and identifying such CNV suppressors could be useful across liquid and solid tumor types.


Assuntos
Leucemia , Neoplasias , Humanos , Pontos de Checagem da Fase M do Ciclo Celular , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , Variações do Número de Cópias de DNA , Heterogeneidade Genética , Aneuploidia , Neoplasias/genética , Neoplasias/metabolismo , Leucemia/genética , Leucemia/metabolismo , Fuso Acromático/metabolismo
2.
J Cell Sci ; 136(11)2023 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-37288769

RESUMO

The mechanical environment of a cell can have many effects, but whether it impacts the DNA sequence of a cell has remained unexamined. To investigate this, we developed a live-cell method to measure changes in chromosome numbers. We edited constitutive genes with GFP or RFP tags on single alleles and discovered that cells that lose Chromosome reporters (ChReporters) become non-fluorescent. We applied our new tools to confined mitosis and to inhibition of the putative tumor suppressor myosin-II. We quantified compression of mitotic chromatin in vivo and demonstrated that similar compression in vitro resulted in cell death, but also rare and heritable ChReptorter loss. Myosin-II suppression rescued lethal multipolar divisions and maximized ChReporter loss during three-dimensional (3D) compression and two-dimensional (2D) lateral confinement, but not in standard 2D culture. ChReporter loss was associated with chromosome mis-segregation, rather than just the number of divisions, and loss in vitro and in mice was selected against in subsequent 2D cultures. Inhibition of the spindle assembly checkpoint (SAC) caused ChReporter loss in 2D culture, as expected, but not during 3D compression, suggesting a SAC perturbation. Thus, ChReporters enable diverse studies of viable genetic changes, and show that confinement and myosin-II affect DNA sequence and mechano-evolution.


Assuntos
Cromossomos , Mitose , Animais , Camundongos , Mitose/genética , Cromossomos/genética , Segregação de Cromossomos/genética , Miosinas/genética , Miosinas/metabolismo , Fuso Acromático/metabolismo , Aneuploidia
3.
Proc Natl Acad Sci U S A ; 118(48)2021 11 30.
Artigo em Inglês | MEDLINE | ID: mdl-34810266

RESUMO

Physicochemical principles such as stoichiometry and fractal assembly can give rise to characteristic scaling between components that potentially include coexpressed transcripts. For key structural factors within the nucleus and extracellular matrix, we discover specific gene-gene scaling exponents across many of the 32 tumor types in The Cancer Genome Atlas, and we demonstrate utility in predicting patient survival as well as scaling-informed machine learning (SIML). All tumors with adjacent tissue data show cancer-elevated proliferation genes, with some genes scaling with the nuclear filament LMNB1, including the transcription factor FOXM1 that we show directly regulates LMNB1 SIML shows that such regulated cancers cluster together with longer overall survival than dysregulated cancers, but high LMNB1 and FOXM1 in half of regulated cancers surprisingly predict poor survival, including for liver cancer. COL1A1 is also studied because it too increases in tumors, and a pan-cancer set of fibrosis genes shows substoichiometric scaling with COL1A1 but predicts patient outcome only for liver cancer-unexpectedly being prosurvival. Single-cell RNA-seq data show nontrivial scaling consistent with power laws from bulk RNA and protein analyses, and SIML segregates synthetic from contractile cancer fibroblasts. Our scaling approach thus yields fundamentals-based power laws relatable to survival, gene function, and experiments.


Assuntos
Fibrose/metabolismo , Lamina Tipo B/química , Neoplasias Hepáticas/metabolismo , Núcleo Celular/metabolismo , Proliferação de Células , Sobrevivência Celular , Colágeno/química , Biologia Computacional , DNA/metabolismo , Matriz Extracelular/metabolismo , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Genômica , Humanos , Estimativa de Kaplan-Meier , Neoplasias Hepáticas/genética , Espectrometria de Massas , Neoplasias/metabolismo , Oncogenes , Prognóstico , Proteômica/métodos , Estresse Mecânico , Transcriptoma , Resultado do Tratamento
4.
Anticancer Drugs ; 34(4): 599-604, 2023 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-36730312

RESUMO

Although Philadelphia chromosome-positive acute leukemia (Ph + -ALL) has been revolutionized with tyrosine kinase inhibitors (TKIs), resistance and mutation are universal events during treatment with first-generation and second-generation TKIs. The present third-generation TKI has a dose-dependent, increased risk of serious cardiovascular events and the sensitivity is poor for patients with ≥2 mutations accompanied by the T315I mutation. Thus, novel and well-tolerated TKIs should be explored. This study analyzes the efficacy and advert effects of olverembatinib, a novel third TKI, in the treatment of newly diagnosed adult Ph + -ALL in induction therapy. Four adult patients with newly diagnosed Ph + -ALL were treated with olverembatinib as the first-line treatment. For induction therapy, these patients received 40 mg of oral olverembatinib quaque omni die for 28 days, 1 mg/kg/d of prednisone for 14 days, then tapered and stopped at 28 days and vindesine 4 mg/d at days 1, 8 and 15. After induction therapy, these patients received median or high-dose of cytarabine and methotrexate combined with oral olverembatinib as consolidation therapy. Then the allogeneic hematopoietic stem cell transplantation (allo-HSCT) was performed. All patients reached complete remission with a complete cytogenetic response after induction therapy. Two patients reached major molecular remission and one with complete molecular remission. Before allo-HSCT, all the patients achieved complete molecular remission. All the patients have survived disease-free for 3-6 months. No severe advert effects were observed. It is well-tolerated and effective for olverembatinib in the treatment of newly diagnosed adult patients with Ph + -ALL. A prospective study should be performed to further testify the role.


Assuntos
Transplante de Células-Tronco Hematopoéticas , Leucemia-Linfoma Linfoblástico de Células Precursoras , Adulto , Humanos , Cromossomo Filadélfia , Estudos Prospectivos , Inibidores de Proteínas Quinases/uso terapêutico , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética
5.
Cell Mol Biol (Noisy-le-grand) ; 69(15): 21-25, 2023 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-38279503

RESUMO

This study was to analyze the correlations of IL-1ß and vitamin D (VitD) with chronic obstructive pulmonary disease assessment test (CAT) score in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). For this purpose, a total of 65 patients with chronic obstructive pulmonary disease (COPD) treated in our hospital between June 2020 and June 2022 were enrolled and assigned to a research group, and 40 healthy individuals who underwent physical examination in our hospital over the same time spanning were enrolled into the control group. The 65 COPD patients were further grouped into a stability group (30 cases) and an exacerbation group (35 cases). The two groups were compared in the levels of 25-hydroxyvitamin D (25(OH)D), interleukin-1ß (IL-1ß) and blood gas indexes (arterial carbon dioxide partial pressure (PaCO2) and arterial partial pressure of oxygen (PaO2). The modified Medical Research Council Dyspnea Scale (mMRC) and the CAT were adopted for evaluation of the stability group and exacerbation group. The correlations of IL-1ß and 25(OH)D with mMRC and CAT scores were analyzed. The diagnostic value of IL-1ß and VitD in patients in different stages was analyzed through receiver operating characteristic (ROC) curves. Results showed that the control group showed greatly lower IL-1ß and PaCO2 levels and higher 25(OH)D and PaO2 levels than the research group (all P<0.05). The stability group got greatly lower mMRC and CAT scores than the exacerbation group (both P<0.05). IL-1ß had positive correlations with mMRC and CAT scores, while 25(OH)D had negative correlations with them (P<0.05). According to ROC curve-based analysis, IL-1ß and 25(OH)D had areas under the curves of 0.814 and 0.583, respectively, in diagnosing the acute exacerbation period, and had specificities of 56.67% and 43.33%, respectively and sensitivities of 97.14% and 74.29%, respectively. In conclusion, patients with COPD have increased IL-1 ß and VitD deficiency, so VitD can be properly supplemented during treatment, and the levels of inflammatory factors should be paid close attention to at all times. IL-1 ß and VitD can be regarded as novel ideas for the diagnosis and treatment of COPD, which may further improve the effect of COPD prevention and treatment.


Assuntos
Doença Pulmonar Obstrutiva Crônica , Humanos , Interleucina-1beta , Oxigênio , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Vitamina D , Vitaminas
6.
Physiol Plant ; 174(3): e13725, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35642076

RESUMO

Rht4 is characterized as a GA-responsive dwarf gene in bread wheat (Triticum aestivum L.). The responsiveness of Rht4 to exogenous GA3 was characterized in seedlings, but the effects of exogenous GA3 on the important morphological and agronomic traits such as plant height, grain-filling rate, and yield components are unclear. In this study, the Rht4 responsiveness of exogenous GA3 on these traits was evaluated using the homozygous F4:5 and F5:6 lines derived from a cross between Jinmai47 and Burt ert937 (Rht4 donor). After exogenous GA3 application, the plant height of the dwarf lines was, on average, increased by 17.54%, about 7.92% more than that of the tall lines. Compared with the tall lines, application of exogenous GA3 significantly increased the kernel weight, maximum grain-filling rate (Gmax), average grain-filling rate (Gave) and kernel weight increment achieving Gmax (Wmax) in both superior and inferior grains, while the day on which the maximum grain-filling rate was reached (Tmax) in Rht4 dwarf lines was significantly earlier in the two generations. What is more, the grain number spike-1 , grain yield plant-1 , and 1000-kernel weight (TKW) of the dwarf lines notably increased after exogenous GA3 -treatment, while there was no significant change in the tall lines except for TKW. The quality traits of the dwarf lines with GA3 -treatment were greatly improved. Taken together, these results suggested that the application of GA3 could improve the grain-filling process of Rht4 and compensate for some negative influences, which may provide a reference for its application in wheat breeding and promote the characterization of its regulatory mechanisms.


Assuntos
Pão , Triticum , Grão Comestível/genética , Fenótipo , Melhoramento Vegetal , Plântula/genética
7.
Luminescence ; 37(9): 1524-1531, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35815832

RESUMO

In this work, a highly efficient electrochemiluminescence (ECL) biosensor was developed based on the nanosponge-hydrogel system for uric acid (UA) detection. First, the nanosponge consisted of polylactic acid glycolic acid (PLGA) nanoparticles immobilized with MoS2 quantum dots (QDs) and urate oxidase (UAO). The marked loading capability of PLGA nanoparticles enables loading many biomolecules and QDs for the specific recognition of UA. Urate oxidase on the nanosponge can catalyze UA to generate H2 O2 in situ, which further triggers the ECL signal for the MoS2 QDs. Furthermore, the biocompatible acrylamide-based hydrogel not only effectively retained the functionalities of the chimeric nanosponge-hydrogel, but also provided structural integrity and engineering flexibility on the electrode for ECL sensing applications. In addition, there were many ester groups and amide bonds in the nanosponge-hydrogel structure. Therefore, many electron can be excited in the ECL process due to the large number of lone electron pairs on oxygen and nitrogen atoms. This resulted in a seven-fold ECL enhancement of the MoS2 QDs. Finally, the nanosponge-hydrogel structure-based ECL biosensor was successfully used in real clinical serum assays. This showed a good analytical performance for UA detection (100-500 µmol/L) with a limit of detection of 20 µmol/L.


Assuntos
Técnicas Biossensoriais , Pontos Quânticos , Técnicas Biossensoriais/métodos , Técnicas Eletroquímicas/métodos , Hidrogéis , Medições Luminescentes/métodos , Molibdênio/química , Pontos Quânticos/química , Urato Oxidase , Ácido Úrico
8.
Molecules ; 27(3)2022 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-35164121

RESUMO

Due to the remarkable anti-tumor activities of oridonin (Ori), research on Rabdosia rubescens has attracted more and more attention in the pharmaceutical field. The purpose of this study was to extract Ori from R. rubescens by ultrasound-assisted extraction (UAE) and prepare Ori liposomes as a novel delivery system to improve the bioavailability and biocompatibility. Response surface methodology (RSM), namely Box-Behnken design (BBD), was applied to optimize extraction conditions, formulation, and preparation process. The results demonstrated that the optimal extraction conditions were an ethanol concentration of 75.9%, an extraction time of 35.7 min, and a solid/liquid ratio of 1:32.6. Under these optimal conditions, the extraction yield of Ori was 4.23 mg/g, which was well matched with the predicted value (4.28 mg/g). The optimal preparation conditions of Ori liposomes by RSM, with an ultrasonic time of 41.1 min, a soybean phospholipids/drug ratio of 9.6 g/g, and a water bath temperature of 53.4 °C, had higher encapsulation efficiency (84.1%). The characterization studies indicated that Ori liposomes had well-dispersible spherical shapes and uniform sizes with a particle size of 137.7 nm, a polydispersity index (PDI) of 0.216, and zeta potential of -24.0 mV. In addition, Ori liposomes presented better activity than free Ori. Therefore, the results indicated that Ori liposomes could enhance the bioactivity of Ori, being proposed as a promising vehicle for drug delivery.


Assuntos
Diterpenos do Tipo Caurano , Isodon/química , Diterpenos do Tipo Caurano/química , Diterpenos do Tipo Caurano/farmacocinética , Diterpenos do Tipo Caurano/farmacologia , Humanos , Lipossomos , Células MCF-7 , Tamanho da Partícula
9.
J Physiol ; 595(23): 7249-7260, 2017 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-28901011

RESUMO

KEY POINTS: In two monogenic models of absence epilepsy, interictal beta/gamma power is augmented in homozygous stargazer (stg/stg) but not homozygous tottering (tg/tg) mice. There are distinct gene-linked patterns of aberrant phase-amplitude coupling in the interictal EEG of both stg/stg and tg/tg mice, compared to +/+ and stg/+ mice. Treatment with ethosuximide significantly blocks seizures in both genotypes, but the abnormal phase-amplitude coupling remains. Seizure-free stg/+ mice have normal power and phase-amplitude coupling, but beta/gamma power is significantly reduced with NMDA receptor blockade, revealing a latent cortical network phenotype that is separable from, and therefore not a result of, seizures. Altogether, these findings reveal gene-linked quantitative electrographic biomarkers free from epileptiform activity, and provide a potential network correlate for persistent cognitive deficits in absence epilepsy despite effective treatment. ABSTRACT: In childhood absence epilepsy, cortical seizures are brief and intermittent; however there are extended periods without behavioural or electrographic ictal events. This genetic disorder is associated with variable degrees of cognitive dysfunction, but no consistent functional biomarkers that might provide insight into interictal cortical function have been described. Previous work in monogenic mouse models of absence epilepsy have shown that the interictal EEG displays augmented beta/gamma power in homozygous stargazer (stg/stg) mice bearing a presynaptic AMPA receptor defect, but not homozygous tottering (tg/tg) mice with a P/Q type calcium channel mutation. To further evaluate the interictal EEG, we quantified phase-amplitude coupling (PAC) in stg/stg, stg/+, tg/tg and wild-type (+/+) mice. We found distinct gene-linked patterns of aberrant PAC in stg/stg and tg/tg mice compared to +/+ and stg/+ mice. Treatment with ethosuximide significantly blocks seizures in both stg/stg and tg/tg, but the abnormal PAC remains. Stg/+ mice are seizure free with normal baseline beta/gamma power and normal theta-gamma PAC, but like stg/stg mice, beta/gamma power is significantly reduced by NMDA receptor blockade, a treatment that paradoxically enhances seizures in stg/stg mice. Stg/+ mice, therefore, have a latent cortical network phenotype that is veiled by NMDA-mediated neurotransmission. Altogether, these findings reveal gene-linked quantitative electrographic biomarkers in the absence of epileptiform activity and provide a potential network correlate for persistent cognitive deficits in absence epilepsy despite effective treatment.


Assuntos
Anticonvulsivantes/farmacologia , Ondas Encefálicas , Epilepsia Tipo Ausência/fisiopatologia , Etossuximida/farmacologia , Genótipo , Animais , Anticonvulsivantes/uso terapêutico , Canais de Cálcio/genética , Canais de Cálcio Tipo N/genética , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/fisiopatologia , Epilepsia Tipo Ausência/tratamento farmacológico , Epilepsia Tipo Ausência/genética , Etossuximida/uso terapêutico , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL
10.
Zhonghua Yi Xue Za Zhi ; 94(29): 2308-11, 2014 Aug 05.
Artigo em Zh | MEDLINE | ID: mdl-25391879

RESUMO

OBJECTIVE: To explore the effect of high resting heart rate (RHR) on the stability of carotid artery plaque in a middle and advanced aged population. METHODS: During 2010-2011, a total of 5 852 participants were selected randomly from 101 510 workers of a Tangshan Kailuan company. RHR was measured and carotid artery ultrasound performed to detect the carotid artery plaque. Logistic regression was performed to analyze the influencing factors of carotid artery stable and unstable plaque. RESULTS: There were 4 894 subjects aged 40-95 years, including 2 945 males and 1 949 females. The detection rates of carotid artery stable and unstable plaque were 25.7% and 25.4% in low RHR group versus 23.8% and 33.2% in high RHR group. Compared with low RHR group, the risk of carotid artery unstable plaque increased to 29% in high RHR group after adjusting for all influencing factors. The odds ratio was 1.290 (95% CI 1.039-1.062). CONCLUSION: High RHR can increase the risk of carotid artery unstable plaque in middle and advanced aged population.


Assuntos
Artérias Carótidas , Estenose das Carótidas/fisiopatologia , Placa Aterosclerótica/fisiopatologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Frequência Cardíaca , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Fatores de Risco
11.
Front Biosci (Landmark Ed) ; 29(2): 62, 2024 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-38420807

RESUMO

BACKGROUND: Mesenchymal cells, including hepatic stellate cells (HSCs), fibroblasts (FBs), myofibroblasts (MFBs), and vascular smooth muscle cells (VSMCs), are the main cells that affect liver fibrosis and play crucial roles in maintaining tissue homeostasis. The dynamic evolution of mesenchymal cells is very important but remains to be explored for researching the reversible mechanism of hepatic fibrosis and its evolution mechanism of hepatic fibrosis to cirrhosis. METHODS: Here, we analysed the transcriptomes of more than 50,000 human single cells from three cirrhotic and three healthy liver tissue samples and the mouse hepatic mesenchymal cells of two healthy and two fibrotic livers to reconstruct the evolutionary trajectory of hepatic mesenchymal cells from a healthy to a cirrhotic state, and a subsequent integrative analysis of bulk RNA sequencing (RNA-seq) data of HSCs from quiescent to active (using transforming growth factor ß1 (TGF-ß1) to stimulate LX-2) to inactive states. RESULTS: We identified core genes and transcription factors (TFs) involved in mesenchymal cell differentiation. In healthy human and mouse livers, the expression of NR1H4 and members of the ZEB families (ZEB1 and ZEB2) changed significantly with the differentiation of FB into HSC and VSMC. In cirrhotic human livers, VSMCs transformed into HSCs with downregulation of MYH11, ACTA2, and JUNB and upregulation of PDGFRB, RGS5, IGFBP5, CD36, A2M, SOX5, and MEF2C. Following HSCs differentiation into MFBs with the upregulation of COL1A1, TIMP1, and NR1H4, a small number of MFBs reverted to inactivated HSCs (iHSCs). The differentiation trajectory of mouse hepatic mesenchymal cells was similar to that in humans; however, the evolution trajectory and proportion of cell subpopulations that reverted from MFBs to iHSCs suggest that the mouse model may not accurately reflect disease progression and outcome in humans. CONCLUSIONS: Our analysis elucidates primary genes and TFs involved in mesenchymal cell differentiation during liver fibrosis using scRNA-seq data, and demonstrated the core genes and TFs in process of HSC activation to MFB and MFB reversal to iHSC using bulk RNA-seq data of human fibrosis induced by TGF-ß1. Furthermore, our findings suggest promising targets for the treatment of liver fibrosis and provide valuable insights into the molecular mechanisms underlying its onset and progression.


Assuntos
Análise da Expressão Gênica de Célula Única , Fatores de Transcrição , Camundongos , Animais , Humanos , Fatores de Transcrição/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Tetracloreto de Carbono/efeitos adversos , Tetracloreto de Carbono/metabolismo , Cirrose Hepática/genética , Cirrose Hepática/metabolismo , Fígado/metabolismo , Diferenciação Celular/genética , Células Estreladas do Fígado/metabolismo
12.
Nat Microbiol ; 9(7): 1752-1763, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38877225

RESUMO

Initiation of development requires differential gene expression and metabolic adaptations. Here we show in the nematode-trapping fungus, Arthrobotrys flagrans, that both are achieved through a dual-function G-protein-coupled receptor (GPCR). A. flagrans develops adhesive traps and recognizes its prey, Caenorhabditis elegans, through nematode-specific pheromones (ascarosides). Gene-expression analyses revealed that ascarosides activate the fungal GPCR, GprC, at the plasma membrane and together with the G-protein alpha subunit GasA, reprograms the cell. However, GprC and GasA also reside in mitochondria and boost respiration. This dual localization of GprC in A. flagrans resembles the localization of the cannabinoid receptor CB1 in humans. The C. elegans ascaroside-sensing GPCR, SRBC66 and GPCRs of many fungi are also predicted for dual localization, suggesting broad evolutionary conservation. An SRBC64/66-GprC chimaeric protein was functional in A. flagrans, and C. elegans SRBC64/66 and DAF38 share ascaroside-binding sites with the fungal GprC receptor, suggesting 400-million-year convergent evolution.


Assuntos
Ascomicetos , Caenorhabditis elegans , Proteínas Fúngicas , Mitocôndrias , Receptores Acoplados a Proteínas G , Animais , Caenorhabditis elegans/microbiologia , Caenorhabditis elegans/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Receptores Acoplados a Proteínas G/genética , Mitocôndrias/metabolismo , Ascomicetos/metabolismo , Ascomicetos/genética , Proteínas Fúngicas/metabolismo , Proteínas Fúngicas/genética , Feromônios/metabolismo , Humanos , Regulação Fúngica da Expressão Gênica
13.
bioRxiv ; 2024 Jan 09.
Artigo em Inglês | MEDLINE | ID: mdl-37066426

RESUMO

Chromosomal instability (CIN), a state in which cells undergo mitotic aberrations that generate chromosome copy number variations, generates aneuploidy and is thought to drive cancer evolution. Although associated with poor prognosis and reduced immune response, CIN generates aneuploidy-induced stresses that could be exploited for immunotherapies. In such contexts, macrophages and the CD47-SIRPα checkpoint are understudied. Here, CIN is induced pharmacologically induced in poorly immunogenic B16F10 mouse melanoma cells, generating persistent micronuclei and diverse aneuploidy while skewing macrophages towards an anti-cancer M1-like phenotype, based on RNA-sequencing profiling, surface marker expression and short-term antitumor studies. These results further translate to in vivo efficacy: Mice bearing CIN-afflicted tumors with wild-type CD47 levels survive only slightly longer relative to chromosomally stable controls, but long-term survival is maximized when combining macrophage-stimulating anti-tumor IgG opsonization and some form of disruption of the CD47-SIRPα checkpoint. Survivors make multi-epitope, de novo anti-cancer IgG that promote macrophage-mediated phagocytosis of CD47 knockout B16F10 cells and suppress tumoroids in vitro and growth of tumors in vivo . CIN does not greatly affect the level of the IgG response compared to previous studies but does significantly increase survival. These results highlight an unexpected therapeutic benefit from CIN when paired with maximal macrophage anti-cancer activity: an anti-cancer vaccination-like antibody response that can lead to more durable cures and further potentiate cell-mediated acquired immunity.

14.
Elife ; 122024 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-38805560

RESUMO

Solid tumors generally exhibit chromosome copy number variation, which is typically caused by chromosomal instability (CIN) in mitosis. The resulting aneuploidy can drive evolution and associates with poor prognosis in various cancer types as well as poor response to T-cell checkpoint blockade in melanoma. Macrophages and the SIRPα-CD47 checkpoint are understudied in such contexts. Here, CIN is induced in poorly immunogenic B16F10 mouse melanoma cells using spindle assembly checkpoint MPS1 inhibitors that generate persistent micronuclei and diverse aneuploidy while skewing macrophages toward a tumoricidal 'M1-like' phenotype based on markers and short-term anti-tumor studies. Mice bearing CIN-afflicted tumors with wild-type CD47 levels succumb similar to controls, but long-term survival is maximized by SIRPα blockade on adoptively transferred myeloid cells plus anti-tumor monoclonal IgG. Such cells are the initiating effector cells, and survivors make de novo anti-cancer IgG that not only promote phagocytosis of CD47-null cells but also suppress tumor growth. CIN does not affect the IgG response, but pairing CIN with maximal macrophage anti-cancer activity increases durable cures that possess a vaccination-like response against recurrence.


Assuntos
Instabilidade Cromossômica , Imunoglobulina G , Macrófagos , Animais , Camundongos , Macrófagos/imunologia , Antígeno CD47/metabolismo , Antígeno CD47/genética , Antígeno CD47/imunologia , Camundongos Endogâmicos C57BL , Melanoma Experimental/imunologia , Melanoma Experimental/terapia , Melanoma Experimental/genética , Linhagem Celular Tumoral , Feminino
18.
Artigo em Inglês | MEDLINE | ID: mdl-36906246

RESUMO

Liver health is important to maintain survival and growth of fish. Currently, the role of dietary docosahexaenoic acid (DHA) in improving fish liver health is largely unknown. This study investigated the role of DHA supplementation in fat deposition and liver damage caused by D-galactosamine (D-GalN) and lipopolysaccharides (LPS) in Nile tilapia (Oreochromis niloticus). Four diets were formulated as control diet (Con), Con supplemented with 1 % DHA, 2 % DHA and 4 % DHA diets, respectively. The diets were fed to 25 Nile tilapia (2.0 ± 0.1 g, average initial weight) in triplicates for four weeks. After the four weeks, 20 fish in each treatment were randomly selected and injected with a mixture of 500 mg D-GalN and 10 µL LPS per mL to induce acute liver injury. The results showed that the Nile tilapia fed on DHA diets decreased visceral somatic index, liver lipid content and serum and liver triglyceride concentrations than those fed on the Con diet. Moreover, after D-GalN/LPS injection, the fish fed on DHA diets decreased alanine aminotransferase and aspartate transaminase activities in the serum. The results of liver qPCR and transcriptomics assays together showed that the DHA diets feeding improved liver health by downregulating the expression of the genes related to toll-like receptor 4 (TLR4) signaling pathway, inflammation and apoptosis. This study indicates that DHA supplementation in Nile tilapia alleviates the liver damage caused by D-GalN/LPS through increasing lipid catabolism, decreasing lipogenesis, TLR4 signaling pathway, inflammation, and apoptosis. Our study provides novel knowledge on the role of DHA in improving liver health in cultured aquatic animals for sustainable aquaculture.


Assuntos
Ciclídeos , Animais , Ração Animal/análise , Ciclídeos/metabolismo , Dieta/veterinária , Suplementos Nutricionais , Ácidos Docosa-Hexaenoicos/farmacologia , Ácidos Docosa-Hexaenoicos/metabolismo , Galactosamina/toxicidade , Galactosamina/metabolismo , Inflamação/metabolismo , Lipopolissacarídeos/toxicidade , Lipopolissacarídeos/metabolismo , Fígado/metabolismo , Receptor 4 Toll-Like/metabolismo
19.
Mol Biol Cell ; 34(13): br19, 2023 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-37903225

RESUMO

Chromosome numbers often change dynamically in tumors and cultured cells, which complicates therapy as well as understanding genotype-mechanotype relationships. Here we use a live-cell "ChReporter" method to identify cells with a single chromosomal loss in efforts to better understand differences in cell shape, motility, and growth. We focus on a standard cancer line and first show clonal populations that retain the ChReporter exhibit large differences in cell and nuclear morphology as well as motility. Phenotype metrics follow simple rules, including migratory persistence scaling with speed, and cytoskeletal differences are evident from drug responses, imaging, and single-cell RNA sequencing. However, mechanotype-genotype relationships between fluorescent ChReporter-positive clones proved complex and motivated comparisons of clones that differ only in loss or retention of a Chromosome-5 ChReporter. When lost, fluorescence-null cells show low expression of Chromosome-5 genes, including a key tumor suppressor APC that regulates microtubules and proliferation. Colonies are compact, nuclei are rounded, and cells proliferate more, with drug results implicating APC, and patient survival data indicating an association in multiple tumor-types. Visual identification of genotype with ChReporters can thus help clarify mechanotype and mechano-evolution.


Assuntos
Aberrações Cromossômicas , Genes Supressores de Tumor , Humanos , Forma Celular , Núcleo Celular , Cromossomos
20.
APL Bioeng ; 6(2): 021504, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35719698

RESUMO

Two meters of DNA in each of our cells must be protected against many types of damage. Mechanoprotection is increasingly understood to be conferred by the nuclear lamina of intermediate filament proteins, but very different patterns of expression and regulation between different cells and tissues remain a challenge to comprehend and translate into applications. We begin with a tutorial style presentation of "tissue blueprints" of lamin expression including single-cell RNA sequencing in major public datasets. Lamin-A, C profiles appear strikingly similar to those for the mechanosensitive factors Vinculin, Yap1, and Piezo1, whereas datasets for lamin-B1 align with and predict regulation by the cell cycle transcription factor, FOXM1, and further predict poor survival across multiple cancers. Various experiments support the distinction between the lamin types and add mechanistic insight into the mechano-regulation of lamin-A, C by both matrix elasticity and externally imposed tissue strain. Both A- and B-type lamins, nonetheless, protect the nucleus from rupture and damage. Ultimately, for mechanically active tissue constructs and organoids as well as cell therapies, lamin levels require particular attention as they help minimize nuclear damage and defects in a cell cycle.

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