tbuckley/cabot-search huggingface dataset:
Noncoding RNAs link metabolic reprogramming to immune microenvironment in cancers
Authors: Yiyin Zhang, Qijiang Mao, Qiming Xia, Jiaxi Cheng, Zhengze Huang, Yirun Li, Peng Chen, Jing Yang, Xiaoxiao Fan, Yuelong Liang, Hui Lin
Journal: Journal of Hematology & Oncology (2021)

Abstract Altered metabolic patterns in tumor cells not only meet their own growth requirements but also shape an immunosuppressive microenvironment through multiple mechanisms. Noncoding RNAs constitute approximately 60% of the transcriptional output of human cells and have been shown to regulate numerous cellular processes under developmental and pathological conditions. Given their extensive action mechanisms based on motif recognition patterns, noncoding RNAs may serve as hinges bridging metabolic activity and immune responses. Indeed, recent studies have shown that microRNAs, long noncoding RNAs and circRNAs are widely involved in tumor metabolic rewiring, immune cell infiltration and function. Hence, we summarized existing knowledge of the role of noncoding RNAs in the remodeling of tumor metabolism and the immune microenvironment, and notably, we established the TIMELnc manual, which is a free and public manual for researchers to identify pivotal lncRNAs that are simultaneously correlated with tumor metabolism and immune cell infiltration based on a bioinformatic approach.
<i>Fusobacterium nucleatum</i> promotes colorectal cancer metastasis through miR-1322/CCL20 axis and M2 polarization
Authors: Chaochao Xu, Lina Fan, Yifeng Lin, Weiyi Shen, Yadong Qi, Ying Zhang, Zhehang Chen, Lan Wang, Yanqin Long, Tongyao Hou, Jianmin Si, Shujie Chen
Journal: Gut Microbes (2021)

Colorectal cancer (CRC) is one of the most common malignant tumors and is associated with Fusobacterium nucleatum (F. nucleatum, Fn) infection. In this study, we explored the role of F. nucleatum in the CRC metastasis. Our results showed that the abundance of F. nucleatum was enriched in the feces and tumors of patients with CRC and tended to increase in stage IV compared to stage I in patients with metastatic CRC. Tumor-derived CCL20 activated by F. nucleatum not only increases CRC metastasis, but also participates in the reprograming of the tumor microenvironment. F. nucleatum promoted macrophage infiltration through CCL20 activation and simultaneously induced M2 macrophage polarization, enhancing the metastasis of CRC. In addition, we identified using database prediction and luciferase activity hat miR-1322, a candidate regulatory micro-RNA, could bind to CCL20 directly. F. nucleatum infection decreased the expression of miR-1322 by activating the NF-κB signaling pathway in CRC cells. In conclusion, F. nucleatum promotes CRC metastasis through the miR-1322/CCL20 axis and M2 polarization.
Tumor microenvironment-based screening repurposes drugs targeting cancer stem cells and cancer-associated fibroblasts
Authors: Pei-Jung Lee, Chao-Chi Ho, Hao Ho, Wan-Jiun Chen, Chiu-Hua Lin, Yi-Hua Lai, Yi-Chen Juan, Wen-Chung Chu, Jia-Hua Lee, Sheng-Fang Su, Hsuan-Yu Chen, Jeremy J. W. Chen, Gee-Chen Chang, Ker-Chau Li, Pan-Chyr Yang, Huei-Wen Chen
Journal: Theranostics (2021)

The tumorous niche may drive the plasticity of heterogeneity and cancer stemness, leading to drug resistance and metastasis, which is the main reason of treatment failure in most cancer patients. The aim of this study was to establish a tumor microenvironment (TME)-based screening to identify drugs that can specifically target cancer stem cells (CSCs) and cancer-associated fibroblasts (CAFs) in the TME. Methods: Lung cancer patient-derived cancer cell and CAFs were utilized to mimic the TME and reproduce the stemness properties of CSCs in vitro and develop a high-throughput drug screening platform with phenotypical parameters. Limiting dilution assay, sphere-forming and ALDH activity assay were utilized to measure the cancer stemness characteristics. In vivo patient-derived xenograft (PDX) models and single-cell RNA sequencing were used to evaluate the mechanisms of the compounds in CSCs and CAFs. Results: The TME-based drug screening platform could comprehensively evaluate the response of cancer cells, CSCs and CAFs to different treatments. Among the 1,524 compounds tested, several drugs were identified to have anti-CAFs, anticancer and anti-CSCs activities. Aloe-emodin and digoxin both show anticancer and anti-CSCs activity in vitro and in vivo, which was further confirmed in the lung cancer PDX model. The combination of digoxin and chemotherapy improved therapeutic efficacy. The single-cell transcriptomics analysis revealed that digoxin could suppress the CSCs subpopulation in CAFs-cocultured cancer cells and cytokine production in CAFs. Conclusions: The TME-based drug screening platform provides a tool to identify and repurpose compounds targeting cancer cells, CSCs and CAFs, which may accelerate drug development and therapeutic application for lung cancer patients.
Sparcl1 promotes nonalcoholic steatohepatitis progression in mice through upregulation of CCL2
Authors: Bin Liu, Liping Xiang, Jing Ji, Wei Liu, Ying Chen, Mingfeng Xia, Yuejun Liu, Wenyue Liu, Peiwu Zhu, Yi Jin, Yu Han, Jieli Lu, Xiaoying Li, Minghua Zheng, Yan Lu
Journal: Journal of Clinical Investigation (2021)

Nonalcoholic fatty liver disease (NAFLD) represents a spectrum of chronic liver disease ranging from simple steatosis (NAFL) to nonalcoholic steatohepatitis (NASH). However, the molecular mechanisms of NASH progression remain incompletely understood. White adipose tissue (WAT) has emerged as an important endocrine organ and contributes not only to the initial stage of NAFLD, but also to its severity. In the current study, through transcriptomic analysis we identified increased expression of Sparcl1, a secreted glycoprotein, in the WAT from NASH mice. Plasma Sparcl1 levels were similarly elevated and positively correlated with hepatic pathological features in NASH patients. Functional studies showed that both chronic injection of recombinant Sparcl1 protein and overexpression of Sparcl1 exaggerated hepatic inflammation and liver injury in mice. In contrast, genetic ablation of Sparcl1, knockdown of Sparcl1 in WAT, and treatment with a Sparcl1-neutralizing antibody dramatically alleviated diet-induced NASH pathogenesis. Mechanistically, Sparcl1 promoted the expression of C-C motif chemokine ligand 2 (CCL2) in hepatocytes through binding to Toll-like receptor 4 (TLR4) and activation of the NF-κB/p65 signaling pathway. Genetically or pharmacologically blocking the CCL2/CCR2 pathway attenuated the hepatic inflammatory response evoked by Sparcl1. Thus, our results demonstrated an important role for Sparcl1 in NASH progression, suggesting a potential target for therapeutic intervention.
Covid-19: Valneva’s vaccine produces stronger immune response than AstraZeneca’s, company reports
Authors: Elisabeth Mahase
Journal: BMJ (2021)

Valneva’s covid-19 vaccine produces significantly higher concentrations of neutralising antibodies than the Oxford-AstraZeneca vaccine does, preliminary trial results show.

The phase III trial included just over 4000 adult volunteers across 26 sites in the UK, of whom 1040 under 30s and 1977 over 30s received Valneva’s vaccine and 995 over 30s were given AstraZeneca’s.

In a press release Valneva reported that the trial met its primary endpoints, with the vaccine showing superiority over the AstraZeneca vaccine in terms of neutralising antibody concentrations and was non-inferior in terms of seroconversion rates (above 95% in both groups), two weeks after the second dose in the over 30s.1

Valneva is a French company that specialises in vaccine development, with …
OA07.03 A Phase II Study of Frontline Rucaparib + Nivolumab in Platinum Sensitive ES SCLC: Interim Analysis
Authors: A. Chauhan, J. Kolesar, D. Yan, A. Maskey, K. Gaurav, R. Mcgarry, J. Miller, A. Khurana, R. O'Neal, J. Veedu, G. Kloecker, C. Park, D. Flora, T. Mullett, M. Evers, L. Anthony, R. Zinner, S. Arnold
Journal: Journal of Thoracic Oncology (2021)
OA04.01 Move For Surgery – A Novel Preconditioning Program to Optimize Health Before Thoracic Surgery: A Randomized Controlled Trial
Authors: Y.S. Patel, I.F. Churchill, K.A. Sullivan, M. Beauchamp, J. Wald, L. Mbuagbaw, J. Agzarian, Y. Shargall, C.J. Finley, C. Fahim, W.C. Hanna
Journal: Journal of Thoracic Oncology (2021)
MA13.08 CHOICE-01: A Phase 3 Study of Toripalimab Versus Placebo in Combination With First-Line Chemotherapy for Advanced NSCLC
Authors: J. Wang, Z. Wang, L. Wu, B. Li, Y. Cheng, X. Li, X. Wang, L. Han, X. Wu, Y. Fan, Y. Yu, D. Lv, J. Shi, J. Huang, S. Zhou, B. Han, G. Sun, Q. Guo, Y. Ji, X. Zhu, S. Hu, W. Zhang, Q. Wang, Y. Jia, Z. Wang, Y. Song, J. Wu, M. Shi, X. Li, Z. Han, Y. Liu, Z. Yu, A. Liu, X. Wang, C. Zhou, D. Zhong, L. Miao, Z. Zhang, H. Zhao, J. Yang, D. Wang, Y. Wang, Q. Li, X. Zhang, M. Ji, Z. Yang, J. Cui, B. Gao, B. Wang, H. Liu, L. Nie, M. He, S. Jin, W. Gu, Y. Shu, T. Zhou, J. Feng, X. Yang, C. Huang, B. Zhu, Y. Yao, Y. Wang, X. Kang, S. Yao, P. Keegan
Journal: Journal of Thoracic Oncology (2021)
OA12.03 Combined Inhibition of SHP2 and CXCR1/2 Promotes Anti-Tumor T Cell Response in NSCLC
Authors: S. Li, K. Tang, A. Khodadadi-Jamayran, J. Jen, H. Han, K. Guidry, T. Chen, Y. Hao, C. Fedele, J. Zebala, D. Maeda, J. Christensen, P. Olson, A. Athanas, K. Wong, B. Neel
Journal: Journal of Thoracic Oncology (2021)
Understanding the impact of the lung microenvironment to enhance the therapeutic potential of mesenchymal stromal cells for acute respiratory distress syndrome
Authors: Claire Masterson, Hector Gonzalez, John G. Laffey
Journal: European Respiratory Journal (2021)

<b>An enhanced understanding of the impact of the lung microenvironment on exogenously administered mesenchymal stromal cells has the potential to enhance their therapeutic potential for ARDS</b>https://bit.ly/2S2fQ1I
MA04.01 Multimodality Treatment and Outcome in Stage III Thymic Epithelial Tumors (TETs): A Retrospective Analysis From the French RYTHMIC Network
Authors: J.C. Benítez, M.V. Bluthgen, M. Boucher, E. Dansin, M. Kerjouan, L. Bigay-Game, E. Pichon, F. Thillays, P. Falcoz, S. Lyubimova, Y. Oulkhouir, F. Calcagno, L. Thiberville, C. Clément-Duchêne, V. Weestel, P. Missy, P. Thomas, J. Maury, T. Molina, N. Girard, B. Besse
Journal: Journal of Thoracic Oncology (2021)
P70.17 Molecular Characteristics and Prognosis TERT Mutations in East Asian Non-Small-Cell Lung Cancer Patients
Authors: W. Gao, C. Xu, W. Wang, D. Wang, Y. Zhu, W. Zhuang, M. Fang, G. Wang, Y. Song, S. Lu, Q. Wang
Journal: Journal of Thoracic Oncology (2021)
P50.04 Amivantamab in Combination With Chemotherapy in Patients With Advanced Non-Small Cell Lung Cancer (NSCLC)
Authors: M. Nagasaka, K. Goto, J. Gomez, T. Hida, C. Shu, C.K. Lee, K. Park, B.C. Cho, J. Lee, S. Ou, C. Bestvina, R. Natale, N. Haddish-Berhane, A. Bhattacharya, R. Verheijen, T. Agrawal, R. Knoblauch, R. Govindan
Journal: Journal of Thoracic Oncology (2021)
Survival Outcomes following LINAC based Stereotactic Radiosurgery/Stereotactic Radiotherapy (SRST) treatment of brain metastases: The Wessex Experience
Authors: Mark Noble, Jeng Ching, Enrico Clarke
Journal: Neuro-Oncology (2021)

Abstract Aims Since 2016, the University Hospital Southampton NHS Foundation Trust (UHSFT) has been commissioned by NHS England to deliver SRST to brain metastases. At UHSFT, all referrals are discussed at the Wessex Neurosciences multidisciplinary team meeting. Referrals that satisfy the criteria set by NHS England (estimated prognosis greater than 6 months, absence or controlled extracranial disease or potentially controllable extracranial disease with a Karnofsky Performance Status &amp;gt;70%) will be offered SRST. This retrospective study was performed to assess overall survival rates of patients with brain metastases treated with SRST with further tumour subtype analysis. We also benchmarked our results with other SRST centres. Method Retrospective data collection was performed for all the patients who have been treated with SRST. Patients who received SRST to a single metastasis, multiple metastases and/or to the resection cavity between 01/01/2017 to 30/09/2019 were included in this study. All treatment was delivered using a LINAC based SRST platform. Prescription doses ranged from 13.5 Gy to 21 Gy in a single fraction, 21 to 24 Gy in 3 fractions and 25 Gy in 5 fractions. Patients are treated using a stereotactic thermoplastic immobilisation shell and dynamic conformal arc therapy with ExacTrac TM and Cone Beam CT imaging. Dates of death were obtained from the NHS Digital Spine and survival analysis using median overall survival was performed using the Kaplan Meier Method. Results 277 patients were treated between 01/01/17 and 30/9/2019. The median overall survival from the Kaplan Meier Method was shown to be 14.7 months and the 6-month overall survival was 71% for all patients. Sub-group analysis of individual tumour sites showed: lung (n=110) median OS 12.1 months, melanoma (n=58) median OS 26.4 months, breast (n=46) median OS not reached (67% still alive) but 18 months survival was 70%, renal (n=22) median OS 15.4 months and colorectal (n=19) median OS 6 months. “Other” tumour sites (n=22) included patients with ovarian, neuroendocrine, sarcoma, testis, oesophagus, unknown primary and gallbladder which were grouped together due to small patient numbers. 41% of patients treated were alive at the time of analysis. Conclusion Patients with brain metastases treated with SRST at UHFST have similar outcomes compared to other SRST centres. These patients have a median overall survival of 14.7 months. However, 29% of patients analysed did not survive more than 6 months. Further collection and analysis of the data might improve patient selection and their outcomes.
Clinical outcomes of relapsed and refractory Hodgkin lymphoma patients after contemporary first-line treatment: a German Hodgkin Study Group analysis
Authors: Paul J. Bröckelmann, Horst Müller, Sarah Gillessen, Xiaoqin Yang, Larissa Koeppel, Veronika Pilz, Patricia Marinello, Peter Kaskel, Monika Raut, Michael Fuchs, Peter Borchmann, Andreas Engert, Bastian von Tresckow
Journal: Leukemia (2021)

Abstract To evaluate patterns of rrHL after contemporary first-line treatment we studied 409 patients with first rrHL (HD13: n = 87, HD14: n = 118, HD15: n = 188, HDR3i: n = 51) at a median age of 37.4 years (18.4–76.8) from the GHSG database. Time to first relapse was ≤12 months in 49% and stage III/IV rrHL present in 52% of patients. In total, 291 patients received high-dose chemotherapy and autologous stem-cell transplantation (ASCT) and intended ASCT failed in 38 patients. ASCT was primarily not intended in 80 patients largely due to low risk disease or age/comorbidities. Overall, 10-year progression-free (PFS) and overall survival (OS) rates after first relapse were 48.2% (95% CI 41.9–54.2%) and 59.4% (95% CI 53.0–65.2%), respectively, with significant differences between subgroups. Inferior survival was observed with no ASCT due to advanced age/comorbidities (five-year PFS 36.2%, 95% CI 17.7–55.0%) or failure of salvage therapy (five-year PFS 36.3%, 95% CI 19.7–53.2%). Similarly, presence of primary refractory disease or stage IV at rrHL conferred inferior survival. In patients with low-risk disease, however, survival appeared favorable even without ASCT (10 y PFS 72.6%, 95% CI 53.7–84.8%). We herein confirm the curative potential of current rrHL treatments providing a robust benchmark to evaluate novel therapeutic strategies in rrHL. Approximately 50% of rrHL patients experienced a consecutive relapse.
The Biosynthesis of Collagen and Its Disorders
Authors: Darwin J. Prockop, Kari I. Kivirikko, Leena Tuderman, Norberto A. Guzman
Journal: New England Journal of Medicine (1979)

(Second of Two Parts)How Do Defects in Biosynthesis Explain Genetic Diseases of Collagen?There are many rare genetic diseases that involve connective tissues, such as skin and bone.As the information reviewed above about collagen structure and biosynthesis accumulated, biochemical data were quickly obtained to explain a number of genetic diseases in man on the basis of molecular defects in collagen biosynthesis. The biochemical data helped to define specific subtypes of some of these diseases. At the same time, study of several of these genetic diseases provided new information about the biosynthetic process itself. For example, shortly after lysyl . . .
Multi-omic evaluation of metabolic alterations in multiple sclerosis identifies shifts in aromatic amino acid metabolism
Authors: Kathryn C. Fitzgerald, Matthew D. Smith, Sol Kim, Elias S. Sotirchos, Michael D. Kornberg, Morgan Douglas, Bardia Nourbakhsh, Jennifer Graves, Ramandeep Rattan, Laila Poisson, Mirela Cerghet, Ellen M. Mowry, Emmanuelle Waubant, Shailendra Giri, Peter A. Calabresi, Pavan Bhargava
Journal: Cell Reports Medicine (2021)

The circulating metabolome provides unique insights into multiple sclerosis (MS) pathophysiology, but existing studies are relatively small or characterized limited metabolites. We test for differences in the metabolome between people with MS (PwMS; n = 637 samples) and healthy controls (HC; n = 317 samples) and assess the association between metabolomic profiles and disability in PwMS. We then assess whether metabolic differences correlate with changes in cellular gene expression using publicly available scRNA-seq data and whether identified metabolites affect human immune cell function. In PwMS, we identify striking abnormalities in aromatic amino acid (AAA) metabolites (p = 2.77E-18) that are also strongly associated with disability (p = 1.01E-4). Analysis of scRNA-seq data demonstrates altered AAA metabolism in CSF and blood-derived monocyte cell populations in PwMS. Treatment with AAA-derived metabolites in vitro alters monocytic endocytosis and pro-inflammatory cytokine production. We identify shifts in AAA metabolism resulting in the reduced production of immunomodulatory metabolites and increased production of metabotoxins in PwMS.
P47.15 A Phase IA Study of Ceritinib + Trametinib in Patients With Advanced ALK- or ROS1- Rearranged NSCLC: Preliminary Results
Authors: M. Lara, M. Gubens, B. Bacaltos, S. Lim, T. Li, D. Gandara, J. Riess, C. Blakely
Journal: Journal of Thoracic Oncology (2021)
Ogfod1 deletion increases cardiac beta-alanine levels and protects mice against ischaemia– reperfusion injury
Authors: Michael Harris, Junhui Sun, Karen Keeran, Angel Aponte, Komudi Singh, Danielle Springer, Marjan Gucek, Mehdi Pirooznia, Matthew E Cockman, Elizabeth Murphy, Leslie M Kennedy
Journal: Cardiovascular Research (2021)

Prolyl hydroxylation is a post-translational modification that regulates protein stability, turnover, and activity. The proteins that catalyze prolyl hydroxylation belong to the 2-oxoglutarate- and iron-dependent oxygenase family of proteins. 2-oxoglutarate- and iron-dependent oxygenase domain-containing protein 1 (Ogfod1), which hydroxylates a proline in ribosomal protein s23 is a newly described member of this family. The aims of this study were to investigate roles for Ogfod1 in the heart, and in the heart's response to stress.We isolated hearts from wild-type (WT) and Ogfod1 knockout (KO) mice and performed quantitative proteomics using tandem mass Tag labelling coupled to liquid chromatography and tandem mass spectrometry (LC-MS/MS) to identify protein changes. Ingenuity pathway analysis identified 'Urate Biosynthesis/Inosine 5'-phosphate Degradation' and 'Purine Nucleotides Degradation II (Aerobic)' as the most significantly enriched pathways. We performed metabolomics analysis and found that both purine and pyrimidine pathways were altered with the purine nucleotide inosine 5'-monophosphate showing a 3.5-fold enrichment in KO hearts (P = 0.011) and the pyrimidine catabolism product beta-alanine showing a 1.7-fold enrichment in KO hearts (P = 0.014). As changes in these pathways have been shown to contribute to cardioprotection, we subjected isolated perfused hearts to ischaemia and reperfusion (I/R). KO hearts showed a 41.4% decrease in infarct size and a 34% improvement in cardiac function compared to WT hearts. This protection was also evident in an in vivo I/R model. Additionally, our data show that treating isolated perfused WT hearts with carnosine, a metabolite of beta-alanine, improved protection in the context of I/R injury, whereas treating KO hearts with carnosine had no impact on recovery of function or infarct size.Taken together, these data show that Ogfod1 deletion alters the myocardial proteome and metabolome to confer protection against I/R injury.
Small extracellular vesicle-mediated targeting of hypothalamic AMPKα1 corrects obesity through BAT activation
Authors: Edward Milbank, Nathalia R. V. Dragano, Ismael González-García, Marcos Rios Garcia, Verónica Rivas-Limeres, Liliana Perdomo, Grégory Hilairet, Francisco Ruiz-Pino, Patricia Mallegol, Donald A. Morgan, Ramón Iglesias-Rey, Cristina Contreras, Luisa Vergori, Juan Cuñarro, Begoña Porteiro, Aleix Gavaldà-Navarro, Rebecca Oelkrug, Anxo Vidal, Juan Roa, Tomás Sobrino, Francesc Villarroya, Carlos Diéguez, Rubén Nogueiras, Cristina García-Cáceres, Manuel Tena-Sempere, Jens Mittag, M. Carmen Martínez, Kamal Rahmouni, Ramaroson Andriantsitohaina, Miguel López
Journal: Nature Metabolism (2021)
Bacteroides vulgatus and Bacteroides dorei predict immune-related adverse events in immune checkpoint blockade treatment of metastatic melanoma
Authors: Mykhaylo Usyk, Abhishek Pandey, Richard B. Hayes, Una Moran, Anna Pavlick, Iman Osman, Jeffrey S. Weber, Jiyoung Ahn
Journal: Genome Medicine (2021)

Abstract Background Immune checkpoint blockade (ICB) shows lasting benefits in advanced melanoma; however, not all patients respond to this treatment and many develop potentially life-threatening immune-related adverse events (irAEs). Identifying individuals who will develop irAEs is critical in order to improve the quality of care. Here, we prospectively demonstrate that the gut microbiome predicts irAEs in melanoma patients undergoing ICB. Methods Pre-, during, and post-treatment stool samples were collected from 27 patients with advanced stage melanoma treated with IPI (anti-CTLA-4) and NIVO (anti-PD1) ICB inhibitors at NYU Langone Health. We completed 16S rRNA gene amplicon sequencing, DNA deep shotgun metagenomic, and RNA-seq metatranscriptomic sequencing. The divisive amplicon denoising algorithm (DADA2) was used to process 16S data. Taxonomy for shotgun sequencing data was assigned using MetaPhlAn2, and gene pathways were assigned using HUMAnN 2.0. Compositionally aware differential expression analysis was performed using ANCOM. The Cox-proportional hazard model was used to assess the prospective role of the gut microbiome (GMB) in irAES, with adjustment for age, sex, BMI, immune ICB treatment type, and sequencing batch. Results Two natural GMB clusters with distinct community compositions were identified from the analysis of 16S rRNA data ( R 2 = 0.16, p &lt; 0.001). In Cox-proportional hazard modeling, these two clusters showed a near 7-fold differential risk for developing irAEs within 1 year of initiating treatment (HR = 6.89 [95% CI: 1.33–35.58]). Using shotgun metagenomics, we further identified 37 bacterial strains differentially expressed between the risk groups, with specific dominance of Bacteroides dorei within the high-risk GMB cluster and Bacteroides vulgatus in the low-risk cluster. The high-risk cluster also appeared to have elevated expression of several functional pathways, including those associated with adenosine metabolism (all FDR &lt; 0.05). A sub-analysis of samples ( n = 10 participants) at baseline and 6 and 12 weeks after the start of treatment revealed that the microbiome remained stable over the course of treatment ( R 2 = 0.88, p &lt; 0.001). Conclusions We identified two distinct fecal bacterial community clusters which are associated differentially with irAEs in ICB-treated advanced melanoma patients.