Retraction for Sorek et al., “Activation Status-Coupled Transient S Acylation Determines Membrane Partitioning of a Plant Rho-Related GTPase”

ABSTRACT ROPs or RACs are plant Rho-related GTPases implicated in the regulation of a multitude of signaling pathways that function at the plasma membrane via posttranslational lipid modifications. The relationships between ROP activation status and membrane localization has not been established. Here, we show that endogenous ROPs, as well as a transgenic His6-green fluorescent protein (GFP)-Arabidopsis thaliana ROP6 (AtROP6) fusion protein, were partitioned between Triton X-100-soluble and -insoluble membranes. In contrast, the His6-GFP-Atrop6CA activated mutant accumulated exclusively in detergent-resistant membranes. GDP induced accumulation of ROPs in Triton-soluble membranes, whereas GTPγS induced accumulation of ROPs in detergent-resistant membranes. Recombinant wild-type and constitutively active AtROP6 proteins were purified from Arabidopsis plants, and in turn, their lipids were cleaved and analyzed by gas chromatography-coupled mass spectrometry. In Triton-soluble membranes, the wild-type AtROP6 was only prenylated, primarily by geranylgeranyl. The activated AtROP6 that accumulated in detergent-resistant membranes was modified by prenyl and acyl lipids, identified as palmitic and stearic acids. Consistently, activated His6-GFP-Atrop6CAmS156, in which C156 was mutated into serine, accumulated in Triton-soluble membranes. These findings show that upon GTP binding and activation, AtROP6, and possibly other ROPs, are transiently S-acylated, inducing their partitioning into detergent-resistant membranes.

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Identification and stratification of systemic lupus erythematosus patients into two transcriptionally distinct clusters based on IFN-I signature

Lupus ◽

10.1177/0961203321990107 ◽

2021 ◽

pp. 096120332199010

Author(s):

Vineeta Shobha ◽

Anu Mohan ◽

AV Malini ◽

Puneet Chopra ◽

Preethi Karunanithi ◽

...

Keyword(s):

Systemic Lupus Erythematosus ◽

Disease Activity ◽

Lupus Erythematosus ◽

Time Course ◽

Cell Activation ◽

Immune Cell ◽

Gene Signature ◽

Systemic Lupus ◽

Auto Antibody ◽

Activation Status

Objective Despite the significant advancement in the understanding of the pathophysiology of systemic lupus erythematosus (SLE) variable clinical response to newer therapies remain a major concern, especially for patients with lupus nephritis and neuropsychiatric systemic lupus erythematosus (NPSLE). We performed this study with an objective to comprehensively characterize Indian SLE patients with renal and neuropsychiatric manifestation with respect to their gene signature, cytokine profile and immune cell phenotypes. Methods We characterized 68 Indian SLE subjects with diverse clinical profiles and disease activity and tried to identify differentially expressed genes and enriched pathways. To understand the temporal profile, same patients were followed at 6 and 12-months intervals. Additionally, auto-antibody profile, levels of various chemokines, cytokines and the proportion of different immune cells and their activation status were captured in these subjects. Results Multiple IFN-related pathways were enriched with significant increase in IFN-I gene signature in SLE patients as compared to normal healthy volunteers (NHV). We identified two transcriptionally distinct clusters within the same cohort of SLE patients with differential immune cell activation status, auto-antibody as well as plasma chemokines and cytokines profile. Conclusions Identification of two distinct clusters of patients based on IFN-I signature provided new insights into the heterogeneity of underlying disease pathogenesis of Indian SLE cohort. Importantly, patient within those clusters retain their distinct expression dynamics of IFN-I signature over the time course of one year despite change in disease activity. This study will guide clinicians and researchers while designing future clinical trials on Indian SLE cohort.

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Optimizing the Procedure to Manufacture Clinical-Grade NK Cells for Adoptive Immunotherapy

Cancers ◽

10.3390/cancers13030577 ◽

2021 ◽

Vol 13 (3) ◽

pp. 577

Author(s):

Adrián Fernández ◽

Alfonso Navarro-Zapata ◽

Adela Escudero ◽

Nerea Matamala ◽

Beatriz Ruz-Caracuel ◽

...

Keyword(s):

Nk Cells ◽

Cell Activation ◽

Mononuclear Cells ◽

Nk Cell ◽

Fold Increase ◽

Clinical Grade ◽

Transcriptome Profile ◽

Expansion Process ◽

Peripheral Blood Mononuclear ◽

Activation Status

Natural killer (NK) cells represent promising tools for cancer immunotherapy. We report the optimization of an NK cell activation–expansion process and its validation on clinical-scale. Methods: RPMI-1640, stem cell growth medium (SCGM), NK MACS and TexMACS were used as culture mediums. Activated and expanded NK cells (NKAE) were obtained by coculturing total peripheral blood mononuclear cells (PBMC) or CD45RA+ cells with irradiated K562mbIL15-41BBL or K562mbIL21-41BBL. Fold increase, NK cell purity, activation status, cytotoxicity and transcriptome profile were analyzed. Clinical-grade NKAE cells were manufactured in CliniMACS Prodigy. Results: NK MACS and TexMACs achieved the highest NK cell purity and lowest T cell contamination. Obtaining NKAE cells from CD45RA+ cells was feasible although PBMC yielded higher total cell numbers and NK cell purity than CD45RA+ cells. The highest fold expansion and NK purity were achieved by using PBMC and K562mbIL21-41BBL cells. However, no differences in activation and cytotoxicity were found when using either NK cell source or activating cell line. Transcriptome profile showed to be different between basal NK cells and NKAE cells expanded with K562mbIL21-41BBL or K562mbIL15-41BBL. Clinical-grade manufactured NKAE cells complied with the specifications from the Spanish Regulatory Agency. Conclusions: GMP-grade NK cells for clinical use can be obtained by using different starting cells and aAPC.

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The role of information structure in the instantiation of objects: Evidence from Amazonian Spanish

Studies in Hispanic and Lusophone Linguistics ◽

10.1515/shll-2020-2028 ◽

2020 ◽

Vol 13 (1) ◽

pp. 219-245

Author(s):

Rosa Vallejos ◽

Evelyn Fernández-Lizárraga ◽

Haley Patterson

Keyword(s):

Regression Analysis ◽

Information Structure ◽

Mixed Model ◽

Linguistic Variables ◽

Social Variables ◽

Null Objects ◽

The Impact ◽

Role Of Information ◽

Activation Status

AbstractThis study analyzes the instantiation of objects in Peruvian Amazonian Spanish (PAS) discourse in two communities with distinct linguistic contexts. We examine the impact of two social variables (gender and place) and nine linguistic variables (transitivity, animacy, definiteness, anaphora function, anaphora expression, cataphora function, cataphora expression, activation, topic persistence) on the speech of eight participants. Our findings indicate that null instantiation in PAS is pervasive, occurring with a range of verb lexemes. While neither gender nor place are significant predictors of null objects, various linguistic variables contribute to the instantiation of objects. The five significant variables as determined by a mixed model regression analysis include the following: animacy, definiteness, anaphora expression, cataphora expression, and activation status. Several findings are consistent with previous research (e. g. human and definite referents disfavor null objects), while other results differ (e. g. PAS propositions disfavor null objects). Activation status and anaphora expression are the most significant predictors of null objects in PAS. In particular, highly accessible referents in discourse and anaphoric null objects favored null objects in subsequent clauses. Thus, the results in the present study demonstrate the pivotal role of information structure in object instantiation, furthering the discussion on syntax-discourse interplay phenomena.

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Peripheral blood natural killer cells activation status determined by CD69 upregulation predicts implantation outcome in IVF

Immunobiology ◽

10.1016/j.imbio.2013.09.002 ◽

2014 ◽

Vol 219 (3) ◽

pp. 167-171 ◽

Cited By ~ 9

Author(s):

Boris V. Dons’koi ◽

Viktor P. Chernyshov ◽

Vera Y. Sirenko ◽

Galina V. Strelko ◽

Dariia V. Osypchuk

Keyword(s):

Natural Killer Cells ◽

Natural Killer ◽

Peripheral Blood ◽

Killer Cells ◽

Activation Status

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Monocyte Functional Responsiveness After PSGL-1–Mediated Platelet Adhesion Is Dependent on Platelet Activation Status

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvbaha.108.167601 ◽

2008 ◽

Vol 28 (8) ◽

pp. 1491-1498 ◽

Cited By ~ 35

Author(s):

Stylianos Bournazos ◽

Jillian Rennie ◽

Simon P. Hart ◽

Keith A.A. Fox ◽

Ian Dransfield

Keyword(s):

Platelet Activation ◽

Platelet Adhesion ◽

Functional Responsiveness ◽

Activation Status

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Mammalian target of rapamycin signaling activation patterns in neuroendocrine tumors of the lung

Endocrine Related Cancer ◽

10.1677/erc-10-0157 ◽

2010 ◽

Vol 17 (4) ◽

pp. 977-987 ◽

Cited By ~ 64

Author(s):

Luisella Righi ◽

Marco Volante ◽

Ida Rapa ◽

Veronica Tavaglione ◽

Frediano Inzani ◽

...

Keyword(s):

Neuroendocrine Tumors ◽

Mammalian Target Of Rapamycin ◽

Mtor Pathway ◽

Differential Sensitivity ◽

Initiation Factor ◽

Target Of Rapamycin ◽

Low Grade ◽

Mtor Inhibition ◽

Signaling Activation ◽

Activation Status

Among alternative therapeutic strategies in clinically aggressive neuroendocrine tumors (NETs) of the lung, promising results have been obtained in experimental clinical trials with mammalian target of rapamycin (mTOR) inhibitors, though in the absence of a proven mTOR signaling activation status. This study analyzed the expression of phosphorylated mTOR (p-mTOR) and its major targets, the ribosomal p70S6-kinase (S6K) and the eukaryotic initiation factor 4E-binding protein 1 (4EBP1) in a large series of 218 surgically resected, malignant lung NETs, including 24 metastasizing typical carcinoids, 73 atypical carcinoids, 60 large cell neuroendocrine carcinomas (LCNECs), and 61 small cell carcinomas (SCLCs). By immunohistochemistry, low-to-intermediate-grade tumors as compared with high-grade tumors showed higher levels of p-mTOR and phosphorylated S6K (p-S6K) (P<0.001), at variance with phosphorylated 4EBP1 (p-4EBP1), which was mainly expressed in LCNECs and SCLCs (P<0.001). The activated status of mTOR pathway was proved by the strong correlation of p-mTOR with p-S6K and somatostatin receptor(s). Western blot analysis of NET tumor samples confirmed such findings, and differential sensitivity to mTOR inhibition according to mTOR pathway activation characteristics was determined in two lung carcinoid cell lines in vitro. None of the investigated molecules had an impact on survival. However, in low-grade tumors, low p-mTOR expression correlated with lymph node metastases (P=0.016), recurrent disease, and survival (P=0.005). In conclusion, these data demonstrate a differential mTOR activation status in the spectrum of pulmonary NETs, possibly suggesting that mTOR pathway profiling might play a predictive role in candidate patients for mTOR-targeted therapies.

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STEM-17. NOT ALL GBM STEM CELLS ARE EQUAL: IMPLICATIONS FOR RESEARCH AND THERAPY

Neuro-Oncology ◽

10.1093/neuonc/noaa215.834 ◽

2020 ◽

Vol 22 (Supplement_2) ◽

pp. ii199-ii200

Author(s):

Luciano Galdieri ◽

Arijita Jash ◽

Olga Malkova ◽

Diane Mao ◽

Jian Campian ◽

...

Keyword(s):

Stem Cells ◽

Phenotypic Diversity ◽

Treatment Resistance ◽

Surface Markers ◽

Mass Cytometry ◽

Pathway Activation ◽

Almost All ◽

And Function ◽

Activation Status

Abstract Glioblastoma (GBM) kills almost all patients within 2 years. A subpopulation of cells, GBM stem cells (GSCs), contributes to treatment resistance and recurrence. A major therapeutic goal is to kill GSCs, but no targeted therapy yet exists. Since their discovery, GSCs have been isolated using single surface markers, such as CD15, CD44, CD133, and a-6 integrin. It remains unknown how these single surface marker-defined GSC populations compare to each other in terms of signal transduction and function and whether expression of different combinations of these markers is associated with distinct phenotypes. Using mass cytometry and fresh operating room specimens, we found that 15 distinct GSC subpopulations exist in vivo and they differ in their MEK/ERK, WNT, and AKT pathway activation status. In culture, some subpopulations were lost and previously undetectable ones materialized. GSCs highly expressing all four surface markers had the greatest self-renewal capacity and in vivo tumorigenicity as well as the strongest WNT pathway activation. This work highlights the signaling and phenotypic diversity in GSC subpopulations, together suggesting that not all GSCs are equivalent. These observations should be considered when studying GSCs in the laboratory, with implications for the development of treatments that target GSCs and prevent tumor recurrence in patients.

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Macrophage activation status rather than repolarization is associated with enhanced checkpoint activity in combination with PI3Kg inhibition

Molecular Cancer Therapeutics ◽

10.1158/1535-7163.mct-20-0961 ◽

2021 ◽

pp. molcanther.0961.2020

Author(s):

Larissa S. Carnevalli ◽

Molly A. Taylor ◽

Matthew King ◽

Anna ML Coenen-Stass ◽

Adina M Hughes ◽

...

Keyword(s):

Macrophage Activation ◽

Activation Status

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The solute carrier MFSD1 decreases β1 integrins activation status and thus tumor metastasis

10.1101/2021.09.17.460743 ◽

2021 ◽

Author(s):

Marko Roblek ◽

Julia Bicher ◽

Merel van Gogh ◽

Attila Gyoergy ◽

Rita Seeboeck ◽

...

Keyword(s):

Tumor Metastasis ◽

Β1 Integrin ◽

Integrin Activation ◽

Tumor Spread ◽

Tumor Cell Migration ◽

Β1 Integrins ◽

Solute Carrier ◽

Patient Prognosis ◽

Turn Over ◽

Activation Status

Solute carriers are increasingly recognized as participating in a plethora of pathologies, including cancer. We describe here the involvement of the orphan solute carrier MFSD1 in the regulation of tumor cell migration. Loss of MFSD1 enabled higher levels of metastasis in a mouse model. We identified an increased migratory potential in MFSD1-/- tumor cells which was mediated by increased focal adhesion turn-over, reduced stability of mature inactive β1 integrin, and the resulting increased integrin activation index. We show that MFSD1 promoted recycling to the cell surface of endocytosed inactive β1 integrin and thereby protected β1 integrin from proteolytic degradation; this led to dampening of the integrin activation index. Furthermore, down-regulation of MFSD1 expression was observed during early steps of tumorigenesis and higher MFSD1 expression levels correlate with a better cancer patient prognosis. In sum, we describe a requirement for endolysosomal MFSD1 in efficient β1 integrin recycling to suppress tumor spread.

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