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1.
Proc Natl Acad Sci U S A ; 120(12): e2219668120, 2023 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-36927156

RESUMO

Root anatomical phenotypes present a promising yet underexploited avenue to deliver major improvements in yield and climate resilience of crops by improving water and nutrient uptake. For instance, the formation of root cortical aerenchyma (RCA) significantly increases soil exploration and resource capture by reducing the metabolic costs of root tissue. A key bottleneck in studying such phenotypes has been the lack of robust high-throughput anatomical phenotyping platforms. We exploited a phenotyping approach based on laser ablation tomography, termed Anatomics, to quantify variation in RCA formation of 436 diverse maize lines in the field. Results revealed a significant and heritable variation for RCA formation. Genome-wide association studies identified a single-nucleotide polymorphism mapping to a root cortex-expressed gene-encoding transcription factor bHLH121. Functional studies identified that the bHLH121 Mu transposon mutant line and CRISPR/Cas9 loss-of-function mutant line showed reduced RCA formation, whereas an overexpression line exhibited significantly greater RCA formation when compared to the wild-type line. Characterization of these lines under suboptimal water and nitrogen availability in multiple soil environments revealed that bHLH121 is required for RCA formation developmentally as well as under studied abiotic stress. Overall functional validation of the bHLH121 gene's importance in RCA formation provides a functional marker to select varieties with improved soil exploration and thus yield under suboptimal conditions.


Assuntos
Fatores de Transcrição , Zea mays , Zea mays/metabolismo , Fatores de Transcrição/metabolismo , Estudo de Associação Genômica Ampla , Raízes de Plantas/metabolismo , Solo , Água/metabolismo
2.
Proc Natl Acad Sci U S A ; 119(17): e2110245119, 2022 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-35446704

RESUMO

Efforts to understand the phenotypic transition that gave rise to maize from teosinte have mainly focused on the analysis of aerial organs, with little insights into possible domestication traits affecting the root system. Archeological excavations in San Marcos cave (Tehuacán, Mexico) yielded two well-preserved 5,300 to 4,970 calibrated y B.P. specimens (SM3 and SM11) corresponding to root stalks composed of at least five nodes with multiple nodal roots and, in case, a complete embryonic root system. To characterize in detail their architecture and anatomy, we used laser ablation tomography to reconstruct a three-dimensional segment of their nodal roots and a scutellar node, revealing exquisite preservation of the inner tissue and cell organization and providing reliable morphometric parameters for cellular characteristics of the stele and cortex. Whereas SM3 showed multiple cortical sclerenchyma typical of extant maize, the scutellar node of the SM11 embryonic root system completely lacked seminal roots, an attribute found in extant teosinte and in two specific maize mutants: root with undetectable meristem1 (rum1) and rootless concerning crown and seminal roots (rtcs). Ancient DNA sequences of SM10­a third San Marcos specimen of equivalent age to SM3 and SM11­revealed the presence of mutations in the transcribed sequence of both genes, offering the possibility for some of these mutations to be involved in the lack of seminal roots of the ancient specimens. Our results indicate that the root system of the earliest maize from Tehuacán resembled teosinte in traits important for maize drought adaptation.


Assuntos
Domesticação , Zea mays , México , Fenótipo , Zea mays/genética
3.
Proc Natl Acad Sci U S A ; 119(30): e2201072119, 2022 07 26.
Artigo em Inglês | MEDLINE | ID: mdl-35858424

RESUMO

Soil compaction represents a major agronomic challenge, inhibiting root elongation and impacting crop yields. Roots use ethylene to sense soil compaction as the restricted air space causes this gaseous signal to accumulate around root tips. Ethylene inhibits root elongation and promotes radial expansion in compacted soil, but its mechanistic basis remains unclear. Here, we report that ethylene promotes abscisic acid (ABA) biosynthesis and cortical cell radial expansion. Rice mutants of ABA biosynthetic genes had attenuated cortical cell radial expansion in compacted soil, leading to better penetration. Soil compaction-induced ethylene also up-regulates the auxin biosynthesis gene OsYUC8. Mutants lacking OsYUC8 are better able to penetrate compacted soil. The auxin influx transporter OsAUX1 is also required to mobilize auxin from the root tip to the elongation zone during a root compaction response. Moreover, osaux1 mutants penetrate compacted soil better than the wild-type roots and do not exhibit cortical cell radial expansion. We conclude that ethylene uses auxin and ABA as downstream signals to modify rice root cell elongation and radial expansion, causing root tips to swell and reducing their ability to penetrate compacted soil.


Assuntos
Ácido Abscísico , Etilenos , Ácidos Indolacéticos , Oryza , Raízes de Plantas , Ácido Abscísico/metabolismo , Etilenos/metabolismo , Ácidos Indolacéticos/metabolismo , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Mutação , Oryza/genética , Oryza/crescimento & desenvolvimento , Oryza/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Solo
4.
J Exp Bot ; 75(18): 5750-5767, 2024 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-38661441

RESUMO

We describe how increased root cortical parenchyma wall width (CPW) can improve tolerance to drought stress in maize by reducing the metabolic costs of soil exploration. Significant variation (1.0-5.0 µm) for CPW was observed in maize germplasm. The functional-structural model RootSlice predicts that increasing CPW from 2 µm to 4 µm is associated with a ~15% reduction in root cortical cytoplasmic volume, respiration rate, and nitrogen content. Analysis of genotypes with contrasting CPW grown with and without water stress in the field confirms that increased CPW is correlated with an ~32-42% decrease in root respiration. Under water stress in the field, increased CPW is correlated with 125% increased stomatal conductance, 325% increased leaf CO2 assimilation rate, 73-78% increased shoot biomass, and 92-108% increased yield. CPW was correlated with leaf mesophyll midrib parenchyma wall width, indicating pleiotropy. Genome-wide association study analysis identified candidate genes underlying CPW. OpenSimRoot modeling predicts that a reduction in root respiration due to increased CPW would also benefit maize growth under suboptimal nitrogen, which requires empirical testing. We propose CPW as a new phene that has utility under edaphic stress meriting further investigation.


Assuntos
Raízes de Plantas , Água , Zea mays , Zea mays/crescimento & desenvolvimento , Zea mays/fisiologia , Zea mays/genética , Zea mays/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/fisiologia , Raízes de Plantas/metabolismo , Água/metabolismo , Secas , Estudo de Associação Genômica Ampla , Desidratação
5.
J Exp Bot ; 75(18): 5989-6005, 2024 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-38970454

RESUMO

Plant phenotypic plasticity plays an important role in nitrogen (N) acquisition and use under nitrogen-limited conditions. However, this role has never been quantified as a function of N availability, leaving it unclear whether plastic responses should be considered as potential targets for selection. A combined modelling and experimentation approach was adopted to quantify the role of plasticity in N uptake and plant yield. Based on a greenhouse experiment we considered plasticity in two maize (Zea mays) traits: root-to-leaf biomass allocation ratio and emergence rate of axial roots. In a simulation experiment we individually enabled or disabled both plastic responses for maize stands grown across six N levels. Both plastic responses contributed to maintaining a higher N uptake, and plant productivity as N availability declined compared with stands in which plastic responses were disabled. We conclude that plastic responses quantified in this study may be a potential target trait in breeding programs for greater N uptake across N levels while it may only be important for the internal use of N under N-limited conditions in maize. Given the complexity of breeding for plastic responses, an a priori model analysis is useful to identify which plastic traits to target for enhanced plant performance.


Assuntos
Modelos Biológicos , Nitrogênio , Raízes de Plantas , Zea mays , Zea mays/crescimento & desenvolvimento , Zea mays/metabolismo , Zea mays/fisiologia , Nitrogênio/metabolismo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Raízes de Plantas/fisiologia , Biomassa , Folhas de Planta/metabolismo , Folhas de Planta/fisiologia , Folhas de Planta/crescimento & desenvolvimento
6.
Proc Natl Acad Sci U S A ; 118(6)2021 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-33536333

RESUMO

Mechanical impedance limits soil exploration and resource capture by plant roots. We examine the role of root anatomy in regulating plant adaptation to mechanical impedance and identify a root anatomical phene in maize (Zea mays) and wheat (Triticum aestivum) associated with penetration of hard soil: Multiseriate cortical sclerenchyma (MCS). We characterize this trait and evaluate the utility of MCS for root penetration in compacted soils. Roots with MCS had a greater cell wall-to-lumen ratio and a distinct UV emission spectrum in outer cortical cells. Genome-wide association mapping revealed that MCS is heritable and genetically controlled. We identified a candidate gene associated with MCS. Across all root classes and nodal positions, maize genotypes with MCS had 13% greater root lignin concentration compared to genotypes without MCS. Genotypes without MCS formed MCS upon exogenous ethylene exposure. Genotypes with MCS had greater lignin concentration and bending strength at the root tip. In controlled environments, MCS in maize and wheat was associated improved root tensile strength and increased penetration ability in compacted soils. Maize genotypes with MCS had root systems with 22% greater depth and 49% greater shoot biomass in compacted soils in the field compared to lines without MCS. Of the lines we assessed, MCS was present in 30 to 50% of modern maize, wheat, and barley cultivars but was absent in teosinte and wild and landrace accessions of wheat and barley. MCS merits investigation as a trait for improving plant performance in maize, wheat, and other grasses under edaphic stress.


Assuntos
Raízes de Plantas/anatomia & histologia , Solo , Triticum/anatomia & histologia , Zea mays/anatomia & histologia , Fenômenos Biomecânicos/efeitos dos fármacos , Etilenos/farmacologia , Genoma de Planta , Estudo de Associação Genômica Ampla , Genótipo , Lignina/metabolismo , Fenótipo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/ultraestrutura , Locos de Características Quantitativas/genética , Espectroscopia de Infravermelho com Transformada de Fourier , Triticum/efeitos dos fármacos , Triticum/genética , Triticum/ultraestrutura , Zea mays/efeitos dos fármacos , Zea mays/genética , Zea mays/ultraestrutura
7.
BMC Plant Biol ; 23(1): 6, 2023 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-36597029

RESUMO

BACKGROUND: Genome wide association (GWA) studies demonstrate linkages between genetic variants and traits of interest. Here, we tested associations between single nucleotide polymorphisms (SNPs) in rice (Oryza sativa) and two root hair traits, root hair length (RHL) and root hair density (RHD). Root hairs are outgrowths of single cells on the root epidermis that aid in nutrient and water acquisition and have also served as a model system to study cell differentiation and tip growth. Using lines from the Rice Diversity Panel-1, we explored the diversity of root hair length and density across four subpopulations of rice (aus, indica, temperate japonica, and tropical japonica). GWA analysis was completed using the high-density rice array (HDRA) and the rice reference panel (RICE-RP) SNP sets. RESULTS: We identified 18 genomic regions related to root hair traits, 14 of which related to RHD and four to RHL. No genomic regions were significantly associated with both traits. Two regions overlapped with previously identified quantitative trait loci (QTL) associated with root hair density in rice. We identified candidate genes in these regions and present those with previously published expression data relevant to root hair development. We re-phenotyped a subset of lines with extreme RHD phenotypes and found that the variation in RHD was due to differences in cell differentiation, not cell size, indicating genes in an associated genomic region may influence root hair cell fate. The candidate genes that we identified showed little overlap with previously characterized genes in rice and Arabidopsis. CONCLUSIONS: Root hair length and density are quantitative traits with complex and independent genetic control in rice. The genomic regions described here could be used as the basis for QTL development and further analysis of the genetic control of root hair length and density. We present a list of candidate genes involved in root hair formation and growth in rice, many of which have not been previously identified as having a relation to root hair growth. Since little is known about root hair growth in grasses, these provide a guide for further research and crop improvement.


Assuntos
Estudo de Associação Genômica Ampla , Oryza , Fenótipo , Locos de Características Quantitativas/genética , Genômica , Diferenciação Celular , Oryza/genética , Polimorfismo de Nucleotídeo Único/genética
8.
Psychother Psychosom ; 92(3): 170-179, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37253335

RESUMO

INTRODUCTION/OBJECTIVE: Treatment results of anorexia nervosa (AN) are modest, with fear of weight gain being a strong predictor of treatment outcome and relapse. Here, we present a virtual reality (VR) setup for exposure to healthy weight and evaluate its potential as an adjunct treatment for AN. METHODS: In two studies, we investigate VR experience and clinical effects of VR exposure to higher weight in 20 women with high weight concern or shape concern and in 20 women with AN. RESULTS: In study 1, 90% of participants (18/20) reported symptoms of high arousal but verbalized low to medium levels of fear. Study 2 demonstrated that VR exposure to healthy weight induced high arousal in patients with AN and yielded a trend that four sessions of exposure improved fear of weight gain. Explorative analyses revealed three clusters of individual reactions to exposure, which need further exploration. CONCLUSIONS: VR exposure is a well-accepted and powerful tool for evoking fear of weight gain in patients with AN. We observed a statistical trend that repeated virtual exposure to healthy weight improved fear of weight gain with large effect sizes. Further studies are needed to determine the mechanisms and differential effects.


Assuntos
Anorexia Nervosa , Realidade Virtual , Humanos , Feminino , Anorexia Nervosa/terapia , Medo , Resultado do Tratamento , Aumento de Peso
9.
Plant Cell Environ ; 45(3): 620-636, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34725839

RESUMO

Mechanical impedance constrains root growth in most soils. Crop cultivation changed the impedance characteristics of native soils, through topsoil erosion, loss of organic matter, disruption of soil structure and loss of biopores. Increasing adoption of Conservation Agriculture in high-input agroecosystems is returning cultivated soils to the soil impedance characteristics of native soils, but in the low-input agroecosystems characteristic of developing nations, ongoing soil degradation is generating more challenging environments for root growth. We propose that root phenotypes have evolved to adapt to the altered impedance characteristics of cultivated soil during crop domestication. The diverging trajectories of soils under Conservation Agriculture and low-input agroecosystems have implications for strategies to develop crops to meet global needs under climate change. We present several root ideotypes as breeding targets under the impedance regimes of both high-input and low-input agroecosystems, as well as a set of root phenotypes that should be useful in both scenarios. We argue that a 'whole plant in whole soil' perspective will be useful in guiding the development of future crops for future soils.


Assuntos
Raízes de Plantas , Solo , Agricultura , Mudança Climática , Produtos Agrícolas
10.
Plant Cell Environ ; 45(3): 789-804, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34453329

RESUMO

Radial expansion is a classic response of roots to a mechanical impedance that has generally been assumed to aid penetration. We analysed the response of maize nodal roots to impedance to test the hypothesis that radial expansion is not related to the ability of roots to cross a compacted soil layer. Genotypes varied in their ability to cross the compacted layer, and those with a steeper approach to the compacted layer or less radial expansion in the compacted layer were more likely to cross the layer and achieve greater depth. Root radial expansion was due to cortical cell size expansion, while cortical cell file number remained constant. Genotypes and nodal root classes that exhibited radial expansion in the compacted soil layer generally also thickened in response to exogenous ethylene in hydroponic culture, that is, radial expansion in response to ethylene was correlated with the thickening response to impedance in soil. We propose that ethylene insensitive roots, that is, those that do not thicken and can overcome impedance, have a competitive advantage under mechanically impeded conditions as they can maintain their elongation rates. We suggest that prolonged exposure to ethylene could function as a stop signal for axial root growth.


Assuntos
Solo , Zea mays , Etilenos , Raízes de Plantas , Zea mays/fisiologia
11.
Plant Cell Environ ; 45(3): 837-853, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34169548

RESUMO

Crops with reduced nutrient and water requirements are urgently needed in global agriculture. Root growth angle plays an important role in nutrient and water acquisition. A maize diversity panel of 481 genotypes was screened for variation in root angle employing a high-throughput field phenotyping platform. Genome-wide association mapping identified several single nucleotide polymorphisms (SNPs) associated with root angle, including one located in the root expressed CBL-interacting serine/threonine-protein kinase 15 (ZmCIPK15) gene (LOC100285495). Reverse genetic studies validated the functional importance of ZmCIPK15, causing a approximately 10° change in root angle in specific nodal positions. A steeper root growth angle improved nitrogen capture in silico and in the field. OpenSimRoot simulations predicted at 40 days of growth that this change in angle would improve nitrogen uptake by 11% and plant biomass by 4% in low nitrogen conditions. In field studies under suboptimal N availability, the cipk15 mutant with steeper growth angles had 18% greater shoot biomass and 29% greater shoot nitrogen accumulation compared to the wild type after 70 days of growth. We propose that a steeper root growth angle modulated by ZmCIPK15 will facilitate efforts to develop new crop varieties with optimal root architecture for improved performance under edaphic stress.


Assuntos
Nitrogênio , Zea mays , Calcineurina/genética , Calcineurina/metabolismo , Estudo de Associação Genômica Ampla , Nitrogênio/metabolismo , Raízes de Plantas/metabolismo , Proteínas Quinases/metabolismo , Serina/genética , Serina/metabolismo , Treonina/metabolismo , Água/metabolismo , Zea mays/metabolismo
12.
Ann Bot ; 130(2): 131-148, 2022 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-35771883

RESUMO

BACKGROUND: Plastic responses of plants to the environment are ubiquitous. Phenotypic plasticity occurs in many forms and at many biological scales, and its adaptive value depends on the specific environment and interactions with other plant traits and organisms. Even though plasticity is the norm rather than the exception, its complex nature has been a challenge in characterizing the expression of plasticity, its adaptive value for fitness and the environmental cues that regulate its expression. SCOPE: This review discusses the characterization and costs of plasticity and approaches, considerations, and promising research directions in studying plasticity. Phenotypic plasticity is genetically controlled and heritable; however, little is known about how organisms perceive, interpret and respond to environmental cues, and the genes and pathways associated with plasticity. Not every genotype is plastic for every trait, and plasticity is not infinite, suggesting trade-offs, costs and limits to expression of plasticity. The timing, specificity and duration of plasticity are critical to their adaptive value for plant fitness. CONCLUSIONS: There are many research opportunities to advance our understanding of plant phenotypic plasticity. New methodology and technological breakthroughs enable the study of phenotypic responses across biological scales and in multiple environments. Understanding the mechanisms of plasticity and how the expression of specific phenotypes influences fitness in many environmental ranges would benefit many areas of plant science ranging from basic research to applied breeding for crop improvement.


Assuntos
Evolução Biológica , Sinais (Psicologia) , Adaptação Fisiológica/genética , Fenótipo , Plantas/genética
13.
Ann Bot ; 130(3): 367-382, 2022 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-35468194

RESUMO

BACKGROUND AND AIMS: Deep roots (i.e. >1 m depth) are important for crops to access water when the topsoil is dry. Root anatomy and hydraulic conductance play important roles in the uptake of soil water, particularly water located deep in the soil. We investigated whether root and xylem anatomy vary as a function of root type, order and length, or with soil depth in roots of two deep-rooted perennial crops: intermediate wheatgrass [Thinopyrum intermedium (Kernza®)] and alfalfa (Medicago sativa). We linked the expression of these anatomical traits to the plants' capacity to take up water from deep soil layers. METHODS: Using laser ablation tomography, we compared the roots of the two crops for cortical area, number and size of metaxylem vessels, and their estimated root axial hydraulic conductance (ERAHCe). The deepest roots investigated were located at soil depths of 2.25 and at 3.5 m in the field and in rhizoboxes, respectively. Anatomical differences were characterized along 1-m-long individual roots, among root types and orders, as well as between environmental conditions. KEY RESULTS: For both crops, a decrease in the number and diameter, or both, of metaxylem vessels along individual root segments and with soil depth in the field resulted in a decrease in ERAHCe. Alfalfa, with a greater number of metaxylem vessels per root throughout the soil profile and, on average, a 4-fold greater ERAHCe, took up more water from the deep soil layers than intermediate wheatgrass. Root anatomical traits were significantly different across root types, classes and growth conditions. CONCLUSIONS: Root anatomical traits are important tools for the selection of crops with enhanced exploitation of deep soil water. The development and breeding of perennial crops for improved subsoil exploitation will be aided by greater understanding of root phenotypes linked to deep root growth and activity.


Assuntos
Medicago sativa , Solo , Produtos Agrícolas/metabolismo , Medicago sativa/metabolismo , Raízes de Plantas , Água/metabolismo , Xilema/metabolismo
14.
Plant Physiol ; 183(3): 1011-1025, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32332090

RESUMO

To test the hypothesis that multiple integrated root phenotypes would co-optimize drought tolerance, we phenotyped the root anatomy and architecture of 400 mature maize (Zea mays) genotypes under well-watered and water-stressed conditions in the field. We found substantial variation in all 23 root phenes measured. A phenotypic bulked segregant analysis revealed that bulks representing the best and worst performers in the field displayed distinct root phenotypes. In contrast to the worst bulk, the root phenotype of the best bulk under drought consisted of greater cortical aerenchyma formation, more numerous and narrower metaxylem vessels, and thicker nodal roots. Partition-against-medians clustering revealed several clusters of unique root phenotypes related to plant performance under water stress. Clusters associated with improved drought tolerance consisted of phene states that likely enable greater soil exploration by reallocating internal resources to greater root construction (increased aerenchyma content, larger cortical cells, fewer cortical cell files), restrict uptake of water to conserve soil moisture (reduced hydraulic conductance, narrow metaxylem vessels), and improve penetrability of hard, dry soils (thick roots with a larger proportion of stele, and smaller distal cortical cells). We propose that the most drought-tolerant-integrated phenotypes merit consideration as breeding ideotypes.


Assuntos
Desidratação/genética , Desidratação/fisiopatologia , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/genética , Raízes de Plantas/fisiologia , Zea mays/genética , Zea mays/fisiologia , Produtos Agrícolas/anatomia & histologia , Produtos Agrícolas/genética , Produtos Agrícolas/fisiologia , Variação Genética , Genótipo , Fenótipo
15.
J Exp Bot ; 71(10): 3185-3197, 2020 05 30.
Artigo em Inglês | MEDLINE | ID: mdl-32080722

RESUMO

Root phenotypes regulate soil resource acquisition; however, their genetic control and phenotypic plasticity are poorly understood. We hypothesized that the responses of root architectural phenes to water deficit (stress plasticity) and different environments (environmental plasticity) are under genetic control and that these loci are distinct. Root architectural phenes were phenotyped in the field using a large maize association panel with and without water deficit stress for three seasons in Arizona and without water deficit stress for four seasons in South Africa. All root phenes were plastic and varied in their plastic response. We identified candidate genes associated with stress and environmental plasticity and candidate genes associated with phenes in well-watered conditions in South Africa and in well-watered and water-stress conditions in Arizona. Few candidate genes for plasticity overlapped with those for phenes expressed under each condition. Our results suggest that phenotypic plasticity is highly quantitative, and plasticity loci are distinct from loci that control phene expression in stress and non-stress, which poses a challenge for breeding programs. To make these loci more accessible to the wider research community, we developed a public online resource that will allow for further experimental validation towards understanding the genetic control underlying phenotypic plasticity.


Assuntos
Raízes de Plantas , Zea mays , Fenótipo , Melhoramento Vegetal , Raízes de Plantas/genética , África do Sul , Zea mays/genética
16.
Biochem J ; 476(16): 2281-2295, 2019 08 22.
Artigo em Inglês | MEDLINE | ID: mdl-31387973

RESUMO

Transmembrane mucin MUC17 is an integral part of the glycocalyx as it covers the brush border membrane of small intestinal enterocytes and presents an extended O-glycosylated mucin domain to the intestinal lumen. Here, we identified two unknown phosphorylated serine residues, S4428 and S4492, in the cytoplasmic tail of human MUC17. We have previously demonstrated that MUC17 is anchored to the apical membrane domain via an interaction with the scaffolding protein PDZK1. S4492, localized in the C-terminal PDZ binding motif of MUC17, was mutated to generate phosphomimetic and phosphodeficient variants of MUC17. Using Caco-2 cells as a model system, we found that induction of an inflammatory state by long-term stimulation with the proinflammatory cytokine TNFα resulted in an increase of MUC17 protein levels and enhanced insertion of MUC17 and its two phospho-variants into apical membranes. Up-regulation and apical insertion of MUC17 was followed by shedding of MUC17-containing vesicles. Transmembrane mucins have previously been shown to play a role in the prevention of bacterial colonization by acting as sheddable decoys for encroaching bacteria. Overexpression and increased presentation at the plasma membrane of wild-type MUC17 and its phosphodeficient variant MUC17 S-4492A protected Caco-2 cells against adhesion of enteropathogenic Escherichia coli, indicating that C-terminal phosphorylation of MUC17 may play a functional role in epithelial cell protection. We propose a new function for MUC17 in inflammation, where MUC17 acts as a second line of defense by preventing attachment of bacteria to the epithelial cell glycocalyx in the small intestine.


Assuntos
Aderência Bacteriana , Escherichia coli Enteropatogênica/metabolismo , Glicocálix/metabolismo , Intestino Delgado/metabolismo , Mucinas/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Regulação para Cima , Substituição de Aminoácidos , Células CACO-2 , Glicocálix/microbiologia , Glicocálix/patologia , Células HEK293 , Humanos , Inflamação/genética , Inflamação/metabolismo , Inflamação/patologia , Intestino Delgado/microbiologia , Intestino Delgado/patologia , Mucinas/genética , Mutação de Sentido Incorreto , Domínios PDZ , Fosforilação/genética , Fator de Necrose Tumoral alfa/genética
17.
Radiologe ; 60(8): 711-720, 2020 Aug.
Artigo em Alemão | MEDLINE | ID: mdl-32710153

RESUMO

BACKGROUND: Cancer immunotherapies play an increasing role in the treatment of advanced cancer. In a subset of patients, atypical response patterns and unconventional adverse events make diagnostic evaluation challenging for radiologists. OBJECTIVES: In this article, we provide a review of the possibilities and limitations of imaging methods in monitoring immunotherapies, discuss the phenomena of pseudoprogression and hyperprogression, and introduce iRECIST as an evaluation standard for clinical studies with immunotherapies. In addition, we describe the most notable adverse events and their imaging features. MATERIALS AND METHODS: This article is based on reviews and studies published since 2009. We used PubMed for the literature search and included the following search terms: "immunotherapy", "checkpoint inhibitor", "pseudoprogression", "iRECIST" and "immune related adverse events". RESULTS AND CONCLUSION: With an incidence of up to 10%, pseudoprogression is a rare phenomenon. Currently, differentiation between pseudoprogression and true progressive disease is only possible by follow-up examinations. iRECIST, published in 2017, introduced immune unconfirmed progressive disease (iUPD) and immune confirmed progressive disease (iCPD) as new categories of therapeutic response. There is still no consensus on the time interval between examinations. Crucial adverse events include hypophysitis and pneumonitis, whereby the latter may present as different patterns of interstitial pneumonia making it difficult to differentiate between drug toxicity, infection, and tumor progression.


Assuntos
Imunoterapia , Neoplasias/terapia , Humanos , Critérios de Avaliação de Resposta em Tumores Sólidos , Resultado do Tratamento
18.
Int J Cancer ; 145(1): 242-253, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-30549269

RESUMO

TG02 is a novel cyclin-dependent kinase (CDK) inhibitor and thought to act mainly via CDK-9 inhibition-dependent depletion of short-lived oncoproteins such as MCL-1 or c-MYC. We studied the activity of TG02 in 9 human long-term glioma cell lines (LTC) and 5 glioma-initiating cell lines (GIC) using various cell death assays in vitro and in the LN-229 LTC and ZH-161 GIC models in vivo. TG02 exhibits strong anti-tumor cell activity with EC50 concentrations in the nanomolar range. Median survival in the LN-229 and ZH-161 models was moderately prolonged by TG02. Neither constitutive CDK levels nor those of MCL-1 or c-MYC correlated with sensitivity to TG02. Cdk-9 or cdk-5 gene silencing alone did not fully reproduce the effects of TG02. C-myc gene silencing inhibited cell growth, but did not modulate TG02 activity. Electron microscopy revealed cell death to be essentially apoptotic. High concentrations of TG02 induced annexin V binding and minor caspase 3 cleavage, but the pan-caspase inhibitor, zVAD-fmk, or BCL-2 or MCL-1 gene transfer only moderately attenuated TG02-induced cell death, and caspase inhibition did not prevent loss of MCL-1 or c-MYC. TG02 activity was independent of O6 -methylguanine DNA methyltransferase expression. Repetitive exposure to TG02 did not generate an acquired TG02 resistance phenotype, but accumulation of MCL-1, loss of c-MYC, or senescence. TG02 is a highly potent apoptosis-inducing agent in glioma cells in vitro. Caspase inhibition does not rescue TG02-treated cells and repetitive exposure fails to confer acquired resistance, supporting the clinical evaluation of TG02 in glioblastoma.


Assuntos
Neoplasias Encefálicas/tratamento farmacológico , Glioblastoma/tratamento farmacológico , Compostos Heterocíclicos de 4 ou mais Anéis/farmacologia , Animais , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Metilases de Modificação do DNA/genética , Metilases de Modificação do DNA/metabolismo , Enzimas Reparadoras do DNA/genética , Enzimas Reparadoras do DNA/metabolismo , Resistencia a Medicamentos Antineoplásicos , Feminino , Expressão Gênica/efeitos dos fármacos , Glioblastoma/genética , Glioblastoma/metabolismo , Glioblastoma/patologia , Compostos Heterocíclicos de 4 ou mais Anéis/farmacocinética , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Inibidores de Proteínas Quinases/farmacologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Distribuição Tecidual , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
19.
Radiology ; 290(2): 290-297, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30422086

RESUMO

Purpose To compare the diagnostic performance of radiomic analysis (RA) and a convolutional neural network (CNN) to radiologists for classification of contrast agent-enhancing lesions as benign or malignant at multiparametric breast MRI. Materials and Methods Between August 2011 and August 2015, 447 patients with 1294 enhancing lesions (787 malignant, 507 benign; median size, 15 mm ± 20) were evaluated. Lesions were manually segmented by one breast radiologist. RA was performed by using L1 regularization and principal component analysis. CNN used a deep residual neural network with 34 layers. All algorithms were also retrained on half the number of lesions (n = 647). Machine interpretations were compared with prospective interpretations by three breast radiologists. Standard of reference was histologic analysis or follow-up. Areas under the receiver operating curve (AUCs) were used to compare diagnostic performance. Results CNN trained on the full cohort was superior to training on the half-size cohort (AUC, 0.88 vs 0.83, respectively; P = .01), but there was no difference for RA and L1 regularization (AUC, 0.81 vs 0.80, respectively; P = .76) or RA and principal component analysis (AUC, 0.78 vs 0.78, respectively; P = .93). By using the full cohort, CNN performance (AUC, 0.88; 95% confidence interval: 0.86, 0.89) was better than RA and L1 regularization (AUC, 0.81; 95% confidence interval: 0.79, 0.83; P < .001) and RA and principal component analysis (AUC, 0.78; 95% confidence interval: 0.76, 0.80; P < .001). However, CNN was inferior to breast radiologist interpretation (AUC, 0.98; 95% confidence interval: 0.96, 0.99; P < .001). Conclusion A convolutional neural network was superior to radiomic analysis for classification of enhancing lesions as benign or malignant at multiparametric breast MRI. Both approaches were inferior to radiologists' performance; however, more training data will further improve performance of convolutional neural network, but not that of radiomics algorithms. © RSNA, 2018 Online supplemental material is available for this article.


Assuntos
Neoplasias da Mama/diagnóstico por imagem , Mama/diagnóstico por imagem , Interpretação de Imagem Assistida por Computador/métodos , Imageamento por Ressonância Magnética/métodos , Redes Neurais de Computação , Feminino , Humanos , Estudos Prospectivos
20.
Plant Cell Environ ; 42(11): 2999-3014, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31314912

RESUMO

Root anatomical phenotypes vary among maize (Zea mays) cultivars and may have adaptive value by modifying the metabolic cost of soil exploration. However, the microbial trade-offs of these phenotypes are unknown. We hypothesized that nodal roots of maize with contrasting cortical anatomy have different patterns of mutualistic and pathogenic fungal colonization. Arbuscular mycorrhizal colonization in the field and mesocosms, root rots in the field, and Fusarium verticillioides colonization in mesocosms were evaluated in maize genotypes with contrasting root cortical anatomy. Increased aerenchyma and decreased living cortical area were associated with decreased mycorrhizal colonization in mesocosm and field experiments with inbred genotypes. In contrast, mycorrhizal colonization of hybrids increased with larger aerenchyma lacunae; this increase coincided with larger root diameters of hybrid roots. F. verticillioides colonization was inversely correlated with living cortical area in mesocosm-grown inbreds, and no relation was found between root rots and living cortical area or aerenchyma in field-grown hybrids. Root rots were positively correlated with cortical cell file number and inversely correlated with cortical cell size. Mycorrhizae and root rots were inversely correlated in field-grown hybrids. We conclude that root anatomy is associated with differential effects on pathogens and mycorrhizal colonization of nodal roots in maize.


Assuntos
Micorrizas/metabolismo , Raízes de Plantas/anatomia & histologia , Zea mays/anatomia & histologia , Zea mays/microbiologia , Fusarium/patogenicidade , Micélio/crescimento & desenvolvimento , Micélio/metabolismo , Micorrizas/crescimento & desenvolvimento , Fenótipo , Doenças das Plantas/microbiologia , Raízes de Plantas/citologia , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/microbiologia , Microbiologia do Solo , Simbiose/fisiologia , Zea mays/crescimento & desenvolvimento , Zea mays/metabolismo
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