Publications by authors named "Young Seok Ju"

72 Publications

Patient-derived organoids as a preclinical platform for precision medicine in colorectal cancer.

Authors:
Young-Won Cho Dong-Wook Min Hwang-Phill Kim Yohan An Sheehyun Kim Jeonghwan Youk Jaeyoung Chun Jong Pil Im Sang-Hyun Song Young Seok Ju Sae-Won Han Kyu Joo Park Tae-You Kim

Mol Oncol 2021 Nov 30. Epub 2021 Nov 30.

Department of Molecular Medicine & Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, Seoul National University, Seoul, Korea.

Patient-derived organoids are being considered as models that can help guide personalized therapy through in vitro anti-cancer drug response evaluation. However, attempts to quantify in vitro drug responses in organoids and compare them with responses in matched patients remain inadequate. In this study, we investigated whether drug responses of organoids correlate with clinical responses of matched patients and disease progression of patients. Organoids were established from 54 patients with colorectal cancer who (except for one patient) did not receive any form of therapy before, and tumor organoids were assessed through whole-exome sequencing. For comparisons of in vitro drug responses in matched patients, we developed an "organoid score" based on the variable anti-cancer treatment responses observed in organoids. Very interestingly, a higher organoid score was significantly correlated with a lower tumor regression rate after the standard-of-care treatment in matched patients. Additionally, we confirmed that patients with a higher organoid score (≥ 2.5) had poorer progression-free survival compared with those with a lower organoid score (< 2.5). Furthermore, to assess potential drug repurposing using an FDA-approved drug library, ten tumor organoids derived from patients with disease progression were applied to a simulation platform. Taken together, organoids and organoid scores can facilitate the prediction of anti-cancer therapy efficacy, and they can be used as a simulation model to determine the next therapeutic options through drug screening. Organoids will be an attractive platform to enable the implementation of personalized therapy for colorectal cancer patients.
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http://dx.doi.org/10.1002/1878-0261.13144DOI Listing
November 2021

Dissecting single-cell genomes through the clonal organoid technique.

Authors:
Jeonghwan Youk Hyun Woo Kwon Ryul Kim Young Seok Ju

Exp Mol Med 2021 Oct 18;53(10):1503-1511. Epub 2021 Oct 18.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 34141, Republic of Korea.

The revolution in genome sequencing technologies has enabled the comprehensive detection of genomic variations in human cells, including inherited germline polymorphisms, de novo mutations, and postzygotic mutations. When these technologies are combined with techniques for isolating and expanding single-cell DNA, the landscape of somatic mosaicism in an individual body can be systematically revealed at a single-cell resolution. Here, we summarize three strategies (whole-genome amplification, microdissection of clonal patches in the tissue, and in vitro clonal expansion of single cells) that are currently applied for single-cell mutational analyses. Among these approaches, in vitro clonal expansion, particularly via adult stem cell-derived organoid culture technologies, yields the most sensitive and precise catalog of somatic mutations in single cells. Moreover, because it produces living mutant cells, downstream validation experiments and multiomics profiling are possible. Through the synergistic combination of organoid culture and genome sequencing, researchers can track genome changes at a single-cell resolution, which will lead to new discoveries that were previously impossible.
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http://dx.doi.org/10.1038/s12276-021-00680-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8569207PMC
October 2021

Severe protein C deficiency in a newborn caused by a homozygous pathogenic variant in the PROC gene: a case report.

Authors:
Uisook Song Young Hye Ryu Kiteak Hong So-Yeon Shim Seongyeol Park Jeong Seok Lee Young Seok Ju Seung Han Shin Soyoung Lee

BMC Pediatr 2021 10 16;21(1):453. Epub 2021 Oct 16.

Department of Pediatrics, Seoul National University Hospital, Seoul National University College of Medicine, 101 Daehak-ro, Jongno-gu, Seoul, 03080, Republic of Korea.

Background: Severe protein C deficiency is a rare and inherited cause of thrombophilia in neonates. Protein C acts as an anticoagulant, and its deficiency results in vascular thrombosis. Herein, we report a case of protein C deficiency with a homozygous pathogenic variant in a term neonate, with good outcomes after proper treatment.

Case Presentation: A four-day-old male newborn was transferred to the Seoul National University Hospital on account of dark red to black skin lesions. He was born full-term with an average birth weight without perinatal problems. There were no abnormal findings in the prenatal tests, including intrauterine sonography. The first skin lesion was observed on his right toes and rapidly progressed to proximal areas, such as the lower legs, left arm, and buttock. Under the impression of thromboembolism or vasculitis, we performed a coagulopathy workup, which revealed a high D-dimer level of 23.05 μg/ml. A skin biopsy showed fibrin clots in most capillaries, and his protein C activity level was below 10%, from which we diagnosed protein C deficiency. On postnatal day 6, he experienced an apnea event with desaturation and an abnormal right pupillary light reflex. Brain computed tomography showed multifocal patchy intracranial hemorrhage and intraventricular hemorrhage with an old ischemic lesion. Ophthalmic examination revealed bilateral retinal traction detachments with retinal folds. Protein C concentrate replacement therapy was added to previous treatments including steroids, prostaglandin E1, and anticoagulation. After replacement therapy, there were no new skin lesions, and the previous lesions recovered with scarring. Although there were no new brain hemorrhagic infarctions, there was ongoing ischemic tissue loss, which required further rehabilitation. Ophthalmic surgical interventions were performed to treat the bilateral retinal traction detachments with retinal folds. Molecular analysis revealed a homozygous pathogenic variant in the PROC gene.

Conclusion: Severe protein C deficiency can manifest as a fatal coagulopathy in any organ. Early diagnosis and proper treatment, including protein C concentrate replacement, may improve outcomes without serious sequelae.
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http://dx.doi.org/10.1186/s12887-021-02923-6DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8520241PMC
October 2021

Mutational spectrum of SARS-CoV-2 during the global pandemic.

Authors:
Kijong Yi Su Yeon Kim Thomas Bleazard Taewoo Kim Jeonghwan Youk Young Seok Ju

Exp Mol Med 2021 08 27;53(8):1229-1237. Epub 2021 Aug 27.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 34141, Korea.

Viruses accumulate mutations under the influence of natural selection and host-virus interactions. Through a systematic comparison of 351,525 full viral genome sequences collected during the recent COVID-19 pandemic, we reveal the spectrum of SARS-CoV-2 mutations. Unlike those of other viruses, the mutational spectrum of SARS-CoV-2 exhibits extreme asymmetry, with a much higher rate of C>U than U>C substitutions, as well as a higher rate of G>U than U>G substitutions. This suggests directional genome sequence evolution during transmission. The substantial asymmetry and directionality of the mutational spectrum enable pseudotemporal tracing of SARS-CoV-2 without prior information about the root sequence, collection time, and sampling region. This shows that the viral genome sequences collected in Asia are similar to the original genome sequence. Adjusted estimation of the dN/dS ratio accounting for the asymmetrical mutational spectrum also shows evidence of negative selection on viral genes, consistent with previous reports. Our findings provide deep insights into the mutational processes in SARS-CoV-2 viral infection and advance the understanding of the history and future evolution of the virus.
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http://dx.doi.org/10.1038/s12276-021-00658-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8393781PMC
August 2021

Clonal dynamics in early human embryogenesis inferred from somatic mutation.

Authors:
Seongyeol Park Nanda Maya Mali Ryul Kim Jeong-Woo Choi Junehawk Lee Joonoh Lim Jung Min Park Jung Woo Park Donghyun Kim Taewoo Kim Kijong Yi June Hyug Choi Seong Gyu Kwon Joo Hee Hong Jeonghwan Youk Yohan An Su Yeon Kim Soo A Oh Youngoh Kwon Dongwan Hong Moonkyu Kim Dong Sun Kim Ji Young Park Ji Won Oh Young Seok Ju

Nature 2021 09 25;597(7876):393-397. Epub 2021 Aug 25.

Graduate School of Medical Science and Engineering (GSMSE), Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.

Cellular dynamics and fate decision in early human embryogenesis remain largely unknown owing to the challenges of performing studies in human embryos. Here, we explored whole-genomes of 334 single-cell colonies and targeted deep sequences of 379 bulk tissues obtained from various anatomical locations of seven recently deceased adult human donors. Using somatic mutations as an intrinsic barcode, we reconstructed early cellular phylogenies that demonstrate (1) an endogenous mutational rate that is higher in the first cell division but decreases to approximately one per cell per cell division later in life; (2) universal unequal contribution of early cells to embryo proper, resulting from early cellular bottlenecks that stochastically set aside epiblast cells within the embryo; (3) examples of varying degrees of early clonal imbalances between tissues on the left and right sides of the body, different germ layers and specific anatomical parts and organs; (4) emergence of a few ancestral cells that will substantially contribute to adult cell pools in blood and liver; and (5) presence of mitochondrial DNA heteroplasmy in the fertilized egg. Our approach also provides insights into the age-related mutational processes and loss of sex chromosomes in normal somatic cells. In sum, this study provides a foundation for future studies to complete cellular phylogenies in human embryogenesis.
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http://dx.doi.org/10.1038/s41586-021-03786-8DOI Listing
September 2021

Single-cell transcriptome of bronchoalveolar lavage fluid reveals sequential change of macrophages during SARS-CoV-2 infection in ferrets.

Authors:
Jeong Seok Lee June-Young Koh Kijong Yi Young-Il Kim Su-Jin Park Eun-Ha Kim Se-Mi Kim Sung Ho Park Young Seok Ju Young Ki Choi Su-Hyung Park

Nat Commun 2021 07 27;12(1):4567. Epub 2021 Jul 27.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.

Few studies have used a longitudinal approach to describe the immune response to SARS-CoV-2 infection. Here, we perform single-cell RNA sequencing of bronchoalveolar lavage fluid cells longitudinally obtained from SARS-CoV-2-infected ferrets. Landscape analysis of the lung immune microenvironment shows distinct changes in cell proportions and characteristics compared to uninfected control, at 2 and 5 days post-infection (dpi). Macrophages are classified into 10 distinct subpopulations with transcriptome changes among monocyte-derived infiltrating macrophages and differentiated M1/M2 macrophages, notably at 2 dpi. Moreover, trajectory analysis reveals gene expression changes from monocyte-derived infiltrating macrophages toward M1 or M2 macrophages and identifies a macrophage subpopulation that has rapidly undergone SARS-CoV-2-mediated activation of inflammatory responses. Finally, we find that M1 or M2 macrophages show distinct patterns of gene modules downregulated by immune-modulatory drugs. Overall, these results elucidate fundamental aspects of the immune response dynamics provoked by SARS-CoV-2 infection.
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http://dx.doi.org/10.1038/s41467-021-24807-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8316405PMC
July 2021

Acquired Resistance to Third-Generation EGFR Tyrosine Kinase Inhibitors in Patients With De Novo EGFR-Mutant NSCLC.

Authors:
Ha-Ram Park Tae Min Kim Yusoo Lee Soyeon Kim Seongyeol Park Young Seok Ju Miso Kim Bhumsuk Keam Yoon Kyung Jeon Dong-Wan Kim Dae Seog Heo

J Thorac Oncol 2021 11 6;16(11):1859-1871. Epub 2021 Jul 6.

Seoul National University Cancer Research Institute, Seoul, Republic of Korea; Department of Internal Medicine, Seoul National University Hospital, Seoul, Republic of Korea.

Introduction: EGFR mostly exists subclonally and is acquired as the most common mechanism of resistance to EGFR tyrosine kinase inhibitors (TKIs). Nevertheless, because de novo EGFR-mutant NSCLC is rare, little is known on acquired resistance mechanisms to third-generation EGFR TKIs.

Methods: Acquired resistance mechanisms were analyzed using tumor and plasma samples before and after third-generation EGFR TKI treatment in four patients with de novo EGFR-mutant NSCLC. Genetic alterations were analyzed by whole-exome sequencing, targeted sequencing, fluorescence in situ hybridization, and droplet digital PCR. MTOR, confirmed for oncogenicity using the Ba/F3 system, was reproduced in H1975 cell lines using CRISPR/Cas9-RNP.

Results: Of seven patients with NSCLC with de novo EGFR mutation, four (LC1-4) who received third-generation EGFR TKIs acquired resistance after achieving a partial response (median = 27 mo, range: 17-48 mo). Novel MTOR and EGFR mutations in cis, MET amplification, and EGFR mutation were identified as acquired resistance mechanisms to third-generation EGFR TKIs. The MTOR mutation was oncogenic in Ba/F3 models and revealed resistance to osimertinib through AKT signaling activation in NCI-H1975 cells harboring the MTOR mutation edited by CRISPR/Cas9 (half-maximal inhibitory concentration, 800 ± 67 nM). Osimertinib in combination with mTOR inhibitors abrogated acquired resistance to osimertinib.

Conclusions: Activation of bypass pathways and the EGFR or EGFR mutation were identified as acquired resistance mechanisms to third-generation EGFR TKIs in patients with NSCLC with de novo EGFR mutation. In addition, MTOR- and EGFR-mutant NSCLC cells were sensitive to osimertinib plus mTOR inhibitors.
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http://dx.doi.org/10.1016/j.jtho.2021.06.013DOI Listing
November 2021

Experimental Models for SARS-CoV-2 Infection.

Authors:
Taewoo Kim Jeong Seok Lee Young Seok Ju

Mol Cells 2021 Jun;44(6):377-383

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea.

Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) is a novel virus that causes coronavirus disease 2019 (COVID-19). To understand the identity, functional characteristics and therapeutic targets of the virus and the diseases, appropriate infection models that recapitulate the pathophysiology of the viral infection are necessary. This article reviews the various infection models, including Vero cells, human cell lines, organoids, and animal models, and discusses their advantages and disadvantages. This knowledge will be helpful for establishing an efficient system for defense against emerging infectious diseases.
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http://dx.doi.org/10.14348/molcells.2021.0094DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8245318PMC
June 2021

Nasal ciliated cells are primary targets for SARS-CoV-2 replication in the early stage of COVID-19.

Authors:
Ji Hoon Ahn JungMo Kim Seon Pyo Hong Sung Yong Choi Myung Jin Yang Young Seok Ju Young Tae Kim Ho Min Kim M D Tazikur Rahman Man Ki Chung Sang Duk Hong Hosung Bae Chang-Seop Lee Gou Young Koh

J Clin Invest 2021 07;131(13)

Center for Vascular Research, Institute for Basic Science (IBS), Daejeon, Republic of Korea.

The upper respiratory tract is compromised in the early period of COVID-19, but SARS-CoV-2 tropism at the cellular level is not fully defined. Unlike recent single-cell RNA-Seq analyses indicating uniformly low mRNA expression of SARS-CoV-2 entry-related host molecules in all nasal epithelial cells, we show that the protein levels are relatively high and that their localizations are restricted to the apical side of multiciliated epithelial cells. In addition, we provide evidence in patients with COVID-19 that SARS-CoV-2 is massively detected and replicated within the multiciliated cells. We observed these findings during the early stage of COVID-19, when infected ciliated cells were rapidly replaced by differentiating precursor cells. Moreover, our analyses revealed that SARS-CoV-2 cellular tropism was restricted to the nasal ciliated versus oral squamous epithelium. These results imply that targeting ciliated cells of the nasal epithelium during the early stage of COVID-19 could be an ideal strategy to prevent SARS-CoV-2 propagation.
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http://dx.doi.org/10.1172/JCI148517DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8245175PMC
July 2021

A large-scale snapshot of intratumor heterogeneity in human cancer.

Authors:
Young Seok Ju

Cancer Cell 2021 04;39(4):463-465

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon 34141, Republic of Korea. Electronic address:

Using sophisticated statistical analyses on population-scale cancer whole-genome sequences, a new study published in Cell characterizes the genomic architecture of intratumor heterogeneity (ITH). It results in an unprecedented snapshot of subclones in about 30 cancer types, generating a wealth of insight into the underlying mutational events, processes, and their selection.
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http://dx.doi.org/10.1016/j.ccell.2021.03.005DOI Listing
April 2021

Cerebral Cavernous Malformation 1 Determines YAP/TAZ Signaling-Dependent Metastatic Hallmarks of Prostate Cancer Cells.

Authors:
Sangryong Park Ho-Young Lee Jayoung Kim Hansol Park Young Seok Ju Eung-Gook Kim Jaehong Kim

Cancers (Basel) 2021 Mar 5;13(5). Epub 2021 Mar 5.

Department of Biochemistry, College of Medicine, Gachon University, Incheon 21999, Korea.

Enhanced Yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ) signaling is correlated with the extraprostatic extension of prostate cancer. However, the mechanism by which YAP/TAZ signaling becomes hyperactive and drives prostate cancer progression is currently unclear. In this study, we revealed that higher expression of CCM1, which is uniquely found in advanced prostate cancer, is inversely correlated with metastasis-free and overall survival in patients with prostate cancer. We also demonstrated that CCM1 induces the metastasis of multiple types of prostate cancer cells by regulating YAP/TAZ signaling. Mechanistically, CCM1, a gene mutated in cerebral cavernous malformation, suppresses DDX5, which regulates the suppression of YAP/TAZ signaling, indicating that CCM1 and DDX5 are novel upstream regulators of YAP/TAZ signaling. Our findings highlight the importance of CCM1-DDX5-YAP/TAZ signaling in the metastasis of prostate cancer cells.
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http://dx.doi.org/10.3390/cancers13051125DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7961486PMC
March 2021

Tumor hypoxia represses γδ T cell-mediated antitumor immunity against brain tumors.

Authors:
Jang Hyun Park Hyun-Jin Kim Chae Won Kim Hyeon Cheol Kim Yujin Jung Hyun-Soo Lee Yunah Lee Young Seok Ju Ji Eun Oh Sung-Hong Park Jeong Ho Lee Sung Ki Lee Heung Kyu Lee

Nat Immunol 2021 03 11;22(3):336-346. Epub 2021 Feb 11.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.

The anatomic location and immunologic characteristics of brain tumors result in strong lymphocyte suppression. Consequently, conventional immunotherapies targeting CD8 T cells are ineffective against brain tumors. Tumor cells escape immunosurveillance by various mechanisms and tumor cell metabolism can affect the metabolic states and functions of tumor-infiltrating lymphocytes. Here, we discovered that brain tumor cells had a particularly high demand for oxygen, which affected γδ T cell-mediated antitumor immune responses but not those of conventional T cells. Specifically, tumor hypoxia activated the γδ T cell protein kinase A pathway at a transcriptional level, resulting in repression of the activatory receptor NKG2D. Alleviating tumor hypoxia reinvigorated NKG2D expression and the antitumor function of γδ T cells. These results reveal a hypoxia-mediated mechanism through which brain tumors and γδ T cells interact and emphasize the importance of γδ T cells for antitumor immunity against brain tumors.
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http://dx.doi.org/10.1038/s41590-020-00860-7DOI Listing
March 2021

Implication of CD69 CD103 tissue-resident-like CD8 T cells as a potential immunotherapeutic target for cholangiocarcinoma.

Authors:
Hyung-Don Kim Seongju Jeong Seongyeol Park Yong Joon Lee Young Seok Ju Danbee Kim Gi-Won Song Jae Hoon Lee Sang-Yeob Kim Jaehoon Shin Eui-Cheol Shin Shin Hwang Changhoon Yoo Su-Hyung Park

Liver Int 2021 04 23;41(4):764-776. Epub 2021 Feb 23.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea.

Background: The heterogeneous immune landscapes of intrahepatic cholangiocarcinoma (ICC) remain largely unknown. Here we aimed to investigate the implications of tissue-resident memory (TRM)-related features of tumour-infiltrating CD8 T cells (CD8 TILs) from ICC patients.

Methods: From ICC patients, we obtained blood samples and ICC surgical specimens (n = 33). We performed multicolour flow cytometry, multiplexed immunohistochemistry and RNA sequencing.

Results: When compared to peripheral CD8 T cells, the CD8 TILs included significantly higher proportions of the CD69 CD103 and CD69 CD103 TRM-like subsets (P < .001 for both). Relative to CD69 and CD69 CD103 cells, the CD69 CD103 CD8 TILs harboured higher levels of T-cell markers representing tumour specificity (ie CD39), proliferation (ie Ki-67) and T-cell activation (ie HLA-DR and CD38) (all P < .001). Moreover, compared to the stroma, the tumour margin and core density each had a significantly higher density of CD103 CD8 TILs (P < .001 for both). ICCs with high proportions of CD69 CD103 cells displayed higher levels of parameters associated with response to immune checkpoint inhibitors (ICIs)-including number of CD8 TIL infiltrates (P = .019), PD-L1 expression in the tumour (P = .046) and expression of the T cell-inflamed gene signature (P < .001). ICCs with lower proportions of CD69 CD103 CD8 TILs exhibited significant enrichment of genes related to the Wnt/β-catenin (P < .001) and TGF-β pathways (P = .002).

Conclusion: CD69 CD103 TRM-like CD8 TILs represent prominent tumour-specific immune responses and hold promise as a potential therapeutic target in ICC patients. Differential TRM-related features of ICCs may help develop future immunotherapeutic strategies such as maximizing TRM responses or inhibiting pathways contributing to immune evasion.
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http://dx.doi.org/10.1111/liv.14814DOI Listing
April 2021

4-1BB co-stimulation further enhances anti-PD-1-mediated reinvigoration of exhausted CD39 CD8 T cells from primary and metastatic sites of epithelial ovarian cancers.

Authors:
Galam Leem Junsik Park Minwoo Jeon Eui-Soon Kim Sang Wun Kim Yong Jae Lee Seong Jin Choi Baekgyu Choi Seongyeol Park Young Seok Ju Inkyung Jung Sunghoon Kim Eui-Cheol Shin Jung Yun Lee Su-Hyung Park

J Immunother Cancer 2020 12;8(2)

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea

Background: Responses to immunotherapy vary between different cancer types and sites. Here, we aimed to investigate features of exhaustion and activation in tumor-infiltrating CD8 T cells at both the primary and metastatic sites in epithelial ovarian cancer.

Methods: Tumor tissues and peripheral blood were obtained from 65 patients with ovarian cancer. From these samples, we isolated tumor-infiltrating lymphocytes (TILs) and peripheral blood mononuclear cells. These cells were used for immunophenotype using multicolor flow cytometry, gene expression profile using RNA sequencing and ex vivo functional restoration assays.

Results: We found that CD39 CD8 TILs were enriched with tumor-specific CD8 TILs, and that the activation status of these cells was determined by the differential programmed cell death protein 1 (PD-1) expression level. CD39 CD8 TILs with high PD-1 expression (PD-1) exhibited features of highly tumor-reactive and terminally exhausted phenotypes. Notably, PD-1 CD39 CD8 TILs showed similar characteristics in terms of T-cell exhaustion and activation between the primary and metastatic sites. Among co-stimulatory receptors, 4-1BB was exclusively overexpressed in CD39 CD8 TILs, especially on PD-1 cells, and 4-1BB-expressing cells displayed immunophenotypes indicating higher degrees of T-cell activation and proliferation, and less exhaustion, compared with cells not expressing 4-1BB. Importantly, 4-1BB agonistic antibodies further enhanced the anti-PD-1-mediated reinvigoration of exhausted CD8 TILs from both primary and metastatic sites.

Conclusion: Severely exhausted PD-1 CD39 CD8 TILs displayed a distinctly heterogeneous exhaustion and activation status determined by differential 4-1BB expression levels, providing rationale and evidence for immunotherapies targeting co-stimulatory receptor 4-1BB in ovarian cancers.
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http://dx.doi.org/10.1136/jitc-2020-001650DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7745695PMC
December 2020

A fusion of CD63-BCAR4 identified in lung adenocarcinoma promotes tumorigenicity and metastasis.

Authors:
Kieun Bae Jin Hee Kim Hyojik Jung Sun-Young Kong Yun-Hee Kim Sunshin Kim Geon Kook Lee Jin Soo Lee Jake June-Koo Lee Young Seok Ju Yang-Kyu Choi Kyong-Ah Yoon

Br J Cancer 2021 01 18;124(1):290-298. Epub 2020 Nov 18.

College of Veterinary Medicine, Konkuk University, Seoul, 05029, Republic of Korea.

Background: Recently, fusion variants of the breast cancer anti-oestrogen-resistance 4 (BCAR4) gene were recurrently discovered in lung adenocarcinoma from the genome-wide studies. However, the functional characterisation of BCAR4 fusion has not been investigated.

Methods: Based on the analysis of RNA-sequencing data, we identified a fusion transcript of CD63-BCAR4 in a Korean patient with lung adenocarcinoma who did not harbour any known activating mutations in EGFR and KRAS genes. To investigate the oncogenic effect of CD63-BCAR4, in vitro and in vivo animal experiments were performed.

Results: In vitro experiments showed strongly enhanced cell migration and proliferation by the exogenous expression of CD63-BCAR4 protein in bronchial epithelial cells. Cell migration was notably reduced after knockdown of BCAR4 fusion by small-interfering RNA. The tumorigenic and metastatic capability of the CD63-BCAR4 fusion was confirmed by using the mouse xenograft model. Fusion-overexpressed cells result in metastasis to the liver and lung as well as the primary tumours after subcutaneous injection into mice. Cyclin D1, MMP1, Slug and mesenchymal markers were significantly increased after CD63-BCAR4 overexpression in the in vitro and in vivo experiments.

Conclusions: Taken together, our results suggest a newly identified fusion gene, CD63-BCAR4 as a potential novel oncogene in lung adenocarcinoma.
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http://dx.doi.org/10.1038/s41416-020-01146-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7782829PMC
January 2021

Three-Dimensional Human Alveolar Stem Cell Culture Models Reveal Infection Response to SARS-CoV-2.

Authors:
Jeonghwan Youk Taewoo Kim Kelly V Evans Young-Il Jeong Yongsuk Hur Seon Pyo Hong Je Hyoung Kim Kijong Yi Su Yeon Kim Kwon Joong Na Thomas Bleazard Ho Min Kim Mick Fellows Krishnaa T Mahbubani Kourosh Saeb-Parsy Seon Young Kim Young Tae Kim Gou Young Koh Byeong-Sun Choi Young Seok Ju Joo-Hyeon Lee

Cell Stem Cell 2020 12 21;27(6):905-919.e10. Epub 2020 Oct 21.

Wellcome-MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge CB2 A0W, UK; Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge CB2 3EL, UK. Electronic address:

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is the cause of a present pandemic, infects human lung alveolar type 2 (hAT2) cells. Characterizing pathogenesis is crucial for developing vaccines and therapeutics. However, the lack of models mirroring the cellular physiology and pathology of hAT2 cells limits the study. Here, we develop a feeder-free, long-term, three-dimensional (3D) culture technique for hAT2 cells derived from primary human lung tissue and investigate infection response to SARS-CoV-2. By imaging-based analysis and single-cell transcriptome profiling, we reveal rapid viral replication and the increased expression of interferon-associated genes and proinflammatory genes in infected hAT2 cells, indicating a robust endogenous innate immune response. Further tracing of viral mutations acquired during transmission identifies full infection of individual cells effectively from a single viral entry. Our study provides deep insights into the pathogenesis of SARS-CoV-2 and the application of defined 3D hAT2 cultures as models for respiratory diseases.
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http://dx.doi.org/10.1016/j.stem.2020.10.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7577700PMC
December 2020

Effects of Cryopreservation and Thawing on Single-Cell Transcriptomes of Human T Cells.

Authors:
Jeong Seok Lee Kijong Yi Young Seok Ju Eui-Cheol Shin

Immune Netw 2020 Aug 15;20(4):e34. Epub 2020 Jul 15.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Korea.

Cryopreservation and thawing of PBMCs are inevitable processes in expanding the scale of experiments in human immunology. Here, we carried out a fundamental study to investigate the detailed effects of PBMC cryopreservation and thawing on transcriptomes. We sorted Tregs from fresh and cryopreserved/thawed PBMCs from an identical donor and performed single-cell RNA-sequencing (scRNA-seq). We found that the cryopreservation and thawing process minimally affects the key molecular features of Tregs, including . However, the cryopreserved and thawed sample had a specific cluster with up-regulation of genes for heat shock proteins. Caution may be warranted in interpreting the character of any cluster of cells with heat shock-related properties when cryopreserved and thawed samples are used for scRNA-seq.
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http://dx.doi.org/10.4110/in.2020.20.e34DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7458795PMC
August 2020

Author Correction: Comprehensive molecular characterization of mitochondrial genomes in human cancers.

Authors:
Yuan Yuan Young Seok Ju Youngwook Kim Jun Li Yumeng Wang Christopher J Yoon Yang Yang Inigo Martincorena Chad J Creighton John N Weinstein Yanxun Xu Leng Han Hyung-Lae Kim Hidewaki Nakagawa Keunchil Park Peter J Campbell Han Liang

Nat Genet 2020 Apr 27. Epub 2020 Apr 27.

Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.

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http://dx.doi.org/10.1038/s41588-020-0629-yDOI Listing
April 2020

The genome-wide landscape of C:G > T:A polymorphism at the CpG contexts in the human population.

Authors:
Jeonghwan Youk Yohan An Seongyeol Park June-Koo Lee Young Seok Ju

BMC Genomics 2020 Mar 30;21(1):270. Epub 2020 Mar 30.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea.

Background: The C:G > T:A substitution at the CpG dinucleotide contexts is the most frequent substitution type in genome evolution. The mutational process is obviously ongoing in the human germline; however, its impact on common and rare genomic polymorphisms has not been comprehensively investigated yet. Here we observed the landscape and dynamics of C:G > T:A substitutions from population-scale human genome sequencing datasets including ~ 4300 whole-genomes from the 1000 Genomes and the pan-cancer analysis of whole genomes (PCAWG) Project and ~ 60,000 whole-exomes from the Exome Aggregation Consortium (ExAC) database.

Results: Of the 28,084,558 CpG sites in the human reference genome, 26.0% show C:G > T:A substitution in the dataset. Remarkably, CpGs in CpG islands (CGIs) have a much lower frequency of such mutations (5.6%). Interestingly, the mutation frequency of CGIs is not uniform with a significantly higher C:G > T:A substitution rate for intragenic CGIs compared to other types. For non-CGI CpGs, the mutation rate was positively correlated with the distance from the nearest CGI up to 2 kb. Finally, we found the impact of negative selection for coding CpG mutations resulting in amino acid change.

Conclusions: This study provides the first unbiased rate of C:G > T:A substitution at the CpG dinucleotide contexts, using population-scale human genome sequencing data. Our findings provide insights into the dynamics of the mutation acquisition in the human genome.
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http://dx.doi.org/10.1186/s12864-020-6674-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7106825PMC
March 2020

Dll4 Suppresses Transcytosis for Arterial Blood-Retinal Barrier Homeostasis.

Authors:
Jee Myung Yang Chan Soon Park Soo Hyun Kim Tae Wook Noh Ju-Hee Kim Seongyeol Park Jingu Lee Jang Ryul Park Dohyun Yoo Hyun Ho Jung Hiroshi Takase David T Shima Markus Schwaninger Seungjoo Lee Il-Kug Kim Junyeop Lee Yong-Sok Ji Sangyong Jon Wang-Yuhl Oh Pilhan Kim Akiyoshi Uemura Young Seok Ju Injune Kim

Circ Res 2020 03 12;126(6):767-783. Epub 2020 Feb 12.

From the Graduate School of Medical Science and Engineering (J.M.Y., C.S.P., S.H.K., T.W.N., J.-H.K., S.P., P.K., Y.S.J., I.K.), Korea Advanced Institute of Science and Technology (KAIST), Daejeon.

Rationale: Central nervous system has low vascular permeability by organizing tight junction (TJ) and limiting endothelial transcytosis. While TJ has long been considered to be responsible for vascular barrier in central nervous system, suppressed transcytosis in endothelial cells is now emerging as a complementary mechanism. Whether transcytosis regulation is independent of TJ and its dysregulation dominantly causes diseases associated with edema remain elusive. Dll4 signaling is important for various vascular contexts, but its role in the maintenance of vascular barrier in central nervous system remains unknown.

Objective: To find a TJ-independent regulatory mechanism selective for transcytosis and identify its dysregulation as a cause of pathological leakage.

Methods And Results: We studied transcytosis in the adult mouse retina with low vascular permeability and employed a hypertension-induced retinal edema model for its pathological implication. Both antibody-based and genetic inactivation of Dll4 or Notch1 induce hyperpermeability by increasing transcytosis without junctional destabilization in arterial endothelial cells, leading to nonhemorrhagic leakage predominantly in the superficial retinal layer. Endothelial deletion represses Dll4 in retinal arteries, phenocopying Dll4 blocking-driven vascular leakage. Ang II (angiotensin II)-induced hypertension represses arterial Sox17 and Dll4, followed by transcytosis-driven retinal edema, which is rescued by a gain of Notch activity. Transcriptomic profiling of retinal endothelial cells suggests that Dll4 blocking activates SREBP1 (sterol regulatory element-binding protein 1)-mediated lipogenic transcription and enriches gene sets favorable for caveolae formation. Profiling also predicts the activation of VEGF (vascular endothelial growth factor) signaling by Dll4 blockade. Inhibition of SREBP1 or VEGF-VEGFR2 (VEGF receptor 2) signaling attenuates both Dll4 blockade-driven and hypertension-induced retinal leakage.

Conclusions: In the retina, Sox17-Dll4-SREBP1 signaling axis controls transcytosis independently of TJ in superficial arteries among heterogeneous regulations for the whole vessels. Uncontrolled transcytosis via dysregulated Dll4 underlies pathological leakage in hypertensive retina and could be a therapeutic target for treating hypertension-associated retinal edema.
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http://dx.doi.org/10.1161/CIRCRESAHA.119.316476DOI Listing
March 2020

Publisher Correction: Comprehensive molecular characterization of mitochondrial genomes in human cancers.

Authors:
Yuan Yuan Young Seok Ju Youngwook Kim Jun Li Yumeng Wang Christopher J Yoon Yang Yang Inigo Martincorena Chad J Creighton John N Weinstein Yanxun Xu Leng Han Hyung-Lae Kim Hidewaki Nakagawa Keunchil Park Peter J Campbell Han Liang

Nat Genet 2020 Feb 11. Epub 2020 Feb 11.

Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.

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http://dx.doi.org/10.1038/s41588-020-0587-4DOI Listing
February 2020

Pan-cancer analysis of whole genomes identifies driver rearrangements promoted by LINE-1 retrotransposition.

Authors:
Bernardo Rodriguez-Martin Eva G Alvarez Adrian Baez-Ortega Jorge Zamora Fran Supek Jonas Demeulemeester Martin Santamarina Young Seok Ju Javier Temes Daniel Garcia-Souto Harald Detering Yilong Li Jorge Rodriguez-Castro Ana Dueso-Barroso Alicia L Bruzos Stefan C Dentro Miguel G Blanco Gianmarco Contino Daniel Ardeljan Marta Tojo Nicola D Roberts Sonia Zumalave Paul A W Edwards Joachim Weischenfeldt Montserrat Puiggròs Zechen Chong Ken Chen Eunjung Alice Lee Jeremiah A Wala Keiran Raine

Nat Genet 2020 03 5;52(3):306-319. Epub 2020 Feb 5.

Cancer Ageing and Somatic Mutation Programme, Wellcome Sanger Institute, Cambridge, UK.

About half of all cancers have somatic integrations of retrotransposons. Here, to characterize their role in oncogenesis, we analyzed the patterns and mechanisms of somatic retrotransposition in 2,954 cancer genomes from 38 histological cancer subtypes within the framework of the Pan-Cancer Analysis of Whole Genomes (PCAWG) project. We identified 19,166 somatically acquired retrotransposition events, which affected 35% of samples and spanned a range of event types. Long interspersed nuclear element (LINE-1; L1 hereafter) insertions emerged as the first most frequent type of somatic structural variation in esophageal adenocarcinoma, and the second most frequent in head-and-neck and colorectal cancers. Aberrant L1 integrations can delete megabase-scale regions of a chromosome, which sometimes leads to the removal of tumor-suppressor genes, and can induce complex translocations and large-scale duplications. Somatic retrotranspositions can also initiate breakage-fusion-bridge cycles, leading to high-level amplification of oncogenes. These observations illuminate a relevant role of L1 retrotransposition in remodeling the cancer genome, with potential implications for the development of human tumors.
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http://dx.doi.org/10.1038/s41588-019-0562-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7058536PMC
March 2020

Comprehensive molecular characterization of mitochondrial genomes in human cancers.

Authors:
Yuan Yuan Young Seok Ju Youngwook Kim Jun Li Yumeng Wang Christopher J Yoon Yang Yang Inigo Martincorena Chad J Creighton John N Weinstein Yanxun Xu Leng Han Hyung-Lae Kim Hidewaki Nakagawa Keunchil Park Peter J Campbell Han Liang

Nat Genet 2020 03 5;52(3):342-352. Epub 2020 Feb 5.

Department of Bioinformatics and Computational Biology, The University of Texas MD Anderson Cancer Center, Houston, TX, USA.

Mitochondria are essential cellular organelles that play critical roles in cancer. Here, as part of the International Cancer Genome Consortium/The Cancer Genome Atlas Pan-Cancer Analysis of Whole Genomes Consortium, which aggregated whole-genome sequencing data from 2,658 cancers across 38 tumor types, we performed a multidimensional, integrated characterization of mitochondrial genomes and related RNA sequencing data. Our analysis presents the most definitive mutational landscape of mitochondrial genomes and identifies several hypermutated cases. Truncating mutations are markedly enriched in kidney, colorectal and thyroid cancers, suggesting oncogenic effects with the activation of signaling pathways. We find frequent somatic nuclear transfers of mitochondrial DNA, some of which disrupt therapeutic target genes. Mitochondrial copy number varies greatly within and across cancers and correlates with clinical variables. Co-expression analysis highlights the function of mitochondrial genes in oxidative phosphorylation, DNA repair and the cell cycle, and shows their connections with clinically actionable genes. Our study lays a foundation for translating mitochondrial biology into clinical applications.
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http://dx.doi.org/10.1038/s41588-019-0557-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7058535PMC
March 2020

PRMT1 Is Required for the Maintenance of Mature β-Cell Identity.

Authors:
Hyunki Kim Byoung-Ha Yoon Chang-Myung Oh Joonyub Lee Kanghoon Lee Heein Song Eunha Kim Kijong Yi Mi-Young Kim Hyeongseok Kim Yong Kyung Kim Eun-Hye Seo Haejeong Heo Hee-Jin Kim Junguee Lee Jae Myoung Suh Seung-Hoi Koo Je Kyung Seong Seyun Kim Young Seok Ju Minho Shong Mirang Kim Hail Kim

Diabetes 2020 03 17;69(3):355-368. Epub 2019 Dec 17.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea

Loss of functional β-cell mass is an essential feature of type 2 diabetes, and maintaining mature β-cell identity is important for preserving a functional β-cell mass. However, it is unclear how β-cells achieve and maintain their mature identity. Here we demonstrate a novel function of protein arginine methyltransferase 1 (PRMT1) in maintaining mature β-cell identity. knockout in fetal and adult β-cells induced diabetes, which was aggravated by high-fat diet-induced metabolic stress. Deletion of in adult β-cells resulted in the immediate loss of histone H4 arginine 3 asymmetric dimethylation (H4R3me2a) and the subsequent loss of β-cell identity. The expression levels of genes involved in mature β-cell function and identity were robustly downregulated as soon as deletion was induced in adult β-cells. Chromatin immunoprecipitation sequencing and assay for transposase-accessible chromatin sequencing analyses revealed that PRMT1-dependent H4R3me2a increases chromatin accessibility at the binding sites for CCCTC-binding factor (CTCF) and β-cell transcription factors. In addition, PRMT1-dependent open chromatin regions may show an association with the risk of diabetes in humans. Together, our results indicate that PRMT1 plays an essential role in maintaining β-cell identity by regulating chromatin accessibility.
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http://dx.doi.org/10.2337/db19-0685DOI Listing
March 2020

FIREVAT: finding reliable variants without artifacts in human cancer samples using etiologically relevant mutational signatures.

Authors:
Hyunbin Kim Andy Jinseok Lee Jongkeun Lee Hyonho Chun Young Seok Ju Dongwan Hong

Genome Med 2019 12 17;11(1):81. Epub 2019 Dec 17.

Bioinformatics Analysis Team, National Cancer Center, 323 Ilsan-ro, Ilsandong-gu, Goyang-si, Gyeonggi-do, 10408, Republic of Korea.

Background: Accurate identification of real somatic variants is a primary part of cancer genome studies and precision oncology. However, artifacts introduced in various steps of sequencing obfuscate confidence in variant calling. Current computational approaches to variant filtering involve intensive interrogation of Binary Alignment Map (BAM) files and require massive computing power, data storage, and manual labor. Recently, mutational signatures associated with sequencing artifacts have been extracted by the Pan-cancer Analysis of Whole Genomes (PCAWG) study. These spectrums can be used to evaluate refinement quality of a given set of somatic mutations.

Results: Here we introduce a novel variant refinement software, FIREVAT (FInding REliable Variants without ArTifacts), which uses known spectrums of sequencing artifacts extracted from one of the largest publicly available catalogs of human tumor samples. FIREVAT performs a quick and efficient variant refinement that accurately removes artifacts and greatly improves the precision and specificity of somatic calls. We validated FIREVAT refinement performance using orthogonal sequencing datasets totaling 384 tumor samples with respect to ground truth. Our novel method achieved the highest level of performance compared to existing filtering approaches. Application of FIREVAT on additional 308 The Cancer Genome Atlas (TCGA) samples demonstrated that FIREVAT refinement leads to identification of more biologically and clinically relevant mutational signatures as well as enrichment of sequence contexts associated with experimental errors. FIREVAT only requires a Variant Call Format file (VCF) and generates a comprehensive report of the variant refinement processes and outcomes for the user.

Conclusions: In summary, FIREVAT facilitates a novel refinement strategy using mutational signatures to distinguish artifactual point mutations called in human cancer samples. We anticipate that FIREVAT results will further contribute to precision oncology efforts that rely on accurate identification of variants, especially in the context of analyzing mutational signatures that bear prognostic and therapeutic significance. FIREVAT is freely available at https://github.com/cgab-ncc/FIREVAT.
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http://dx.doi.org/10.1186/s13073-019-0695-xDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6916105PMC
December 2019

Germline gain-of-function mutation of STAT1 rescued by somatic mosaicism in immune dysregulation-polyendocrinopathy-enteropathy-X-linked-like disorder.

Authors:
Jeong Seok Lee Yohan An Christopher J Yoon Jeong Yeon Kim Kyung Hwan Kim Alexandra F Freeman Jae-Joon Yim Eui-Cheol Shin Steven M Holland Eun Young Lee Young Seok Ju

J Allergy Clin Immunol 2020 03 2;145(3):1017-1021. Epub 2019 Dec 2.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Korea; Biomedical Science and Engineering Interdisciplinary Program, Korea Advanced Institute of Science and Technology, Korea. Electronic address:

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http://dx.doi.org/10.1016/j.jaci.2019.11.028DOI Listing
March 2020

4-1BB Delineates Distinct Activation Status of Exhausted Tumor-Infiltrating CD8 T Cells in Hepatocellular Carcinoma.

Authors:
Hyung-Don Kim Seongyeol Park Seongju Jeong Yong Joon Lee Hoyoung Lee Chang Gon Kim Kyung Hwan Kim Seung-Mo Hong Jung-Yun Lee Sunghoon Kim Hong Kwan Kim Byung Soh Min Jong Hee Chang Young Seok Ju Eui-Cheol Shin Gi-Won Song Shin Hwang Su-Hyung Park

Hepatology 2020 03 18;71(3):955-971. Epub 2019 Oct 18.

Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology, Daejeon, Republic of Korea.

Background And Aims: Targeting costimulatory receptors with agonistic antibodies is a promising cancer immunotherapy option. We aimed to investigate costimulatory receptor expression, particularly 4-1BB (CD137 or tumor necrosis factor receptor superfamily member 9), on tumor-infiltrating CD8 T cells (CD8 tumor-infiltrating lymphocytes [TILs]) and its association with distinct T-cell activation features among exhausted CD8 TILs in hepatocellular carcinoma (HCC).

Approach And Results: Tumor tissues, adjacent nontumor tissues, and peripheral blood were collected from HCC patients undergoing surgical resection (n = 79). Lymphocytes were isolated and used for multicolor flow cytometry, RNA-sequencing, and in vitro functional restoration assays. Among the examined costimulatory receptors, 4-1BB was most prominently expressed on CD8 TILs. 4-1BB expression was almost exclusively detected on CD8 T cells in the tumor-especially on programmed death 1 (PD-1) cells and not PD-1 and PD-1 cells. Compared to PD-1 and 4-1BB PD-1 CD8 TILs, 4-1BB PD-1 CD8 TILs exhibited higher levels of tumor reactivity and T-cell activation markers and significant enrichment for T-cell activation gene signatures. Per-patient analysis revealed positive correlations between percentages of 4-1BB cells among CD8 TILs and levels of parameters of tumor reactivity and T-cell activation. Among highly exhausted PD-1 CD8 TILs, 4-1BB cells harbored higher proportions of cells with proliferative and reinvigoration potential. Our 4-1BB-related gene signature predicted survival outcomes of HCC patients in the The Cancer Genome Atlas cohort. 4-1BB agonistic antibodies enhanced the function of CD8 TILs and further enhanced the anti-PD-1-mediated reinvigoration of CD8 TILs, especially in cases showing high levels of T-cell activation.

Conclusion: 4-1BB expression on CD8 TILs represents a distinct activation state among highly exhausted CD8 T cells in HCC. 4-1BB costimulation with agonistic antibodies may be a promising strategy for treating HCCs exhibiting prominent T-cell activation.
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http://dx.doi.org/10.1002/hep.30881DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7154753PMC
March 2020

Tracing Oncogene Rearrangements in the Mutational History of Lung Adenocarcinoma.

Authors:
Jake June-Koo Lee Seongyeol Park Hansol Park Sehui Kim Jongkeun Lee Junehawk Lee Jeonghwan Youk Kijong Yi Yohan An In Kyu Park Chang Hyun Kang Doo Hyun Chung Tae Min Kim Yoon Kyung Jeon Dongwan Hong Peter J Park Young Seok Ju Young Tae Kim

Cell 2019 06 30;177(7):1842-1857.e21. Epub 2019 May 30.

Department of Thoracic and Cardiovascular Surgery, Seoul National University Hospital, Seoul 03080, Korea; Seoul National University Cancer Research Institute, Seoul 03080, Korea. Electronic address:

Mutational processes giving rise to lung adenocarcinomas (LADCs) in non-smokers remain elusive. We analyzed 138 LADC whole genomes, including 83 cases with minimal contribution of smoking-associated mutational signature. Genomic rearrangements were not correlated with smoking-associated mutations and frequently served as driver events of smoking-signature-low LADCs. Complex genomic rearrangements, including chromothripsis and chromoplexy, generated 74% of known fusion oncogenes, including EML4-ALK, CD74-ROS1, and KIF5B-RET. Unlike other collateral rearrangements, these fusion-oncogene-associated rearrangements were frequently copy-number-balanced, representing a genomic signature of early oncogenesis. Analysis of mutation timing revealed that fusions and point mutations of canonical oncogenes were often acquired in the early decades of life. During a long latency, cancer-related genes were disrupted or amplified by complex rearrangements. The genomic landscape was different between subgroups-EGFR-mutant LADCs had frequent whole-genome duplications with p53 mutations, whereas fusion-oncogene-driven LADCs had frequent SETD2 mutations. Our study highlights LADC oncogenesis driven by endogenous mutational processes.
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http://dx.doi.org/10.1016/j.cell.2019.05.013DOI Listing
June 2019

Characterizing Mutational Signatures in Human Cancer Cell Lines Reveals Episodic APOBEC Mutagenesis.

Authors:
Mia Petljak Ludmil B Alexandrov Jonathan S Brammeld Stacey Price David C Wedge Sebastian Grossmann Kevin J Dawson Young Seok Ju Francesco Iorio Jose M C Tubio Ching Chiek Koh Ilias Georgakopoulos-Soares Bernardo Rodríguez-Martín Burçak Otlu Sarah O'Meara Adam P Butler Andrew Menzies Shriram G Bhosle Keiran Raine David R Jones Jon W Teague Kathryn Beal Calli Latimer Laura O'Neill Jorge Zamora Elizabeth Anderson Nikita Patel Mark Maddison Bee Ling Ng Jennifer Graham

Cell 2019 03;176(6):1282-1294.e20

Cytometry Core Facility, Wellcome Sanger Institute, Hinxton, Cambridgeshire CB10 1SA, UK.

Multiple signatures of somatic mutations have been identified in cancer genomes. Exome sequences of 1,001 human cancer cell lines and 577 xenografts revealed most common mutational signatures, indicating past activity of the underlying processes, usually in appropriate cancer types. To investigate ongoing patterns of mutational-signature generation, cell lines were cultured for extended periods and subsequently DNA sequenced. Signatures of discontinued exposures, including tobacco smoke and ultraviolet light, were not generated in vitro. Signatures of normal and defective DNA repair and replication continued to be generated at roughly stable mutation rates. Signatures of APOBEC cytidine deaminase DNA-editing exhibited substantial fluctuations in mutation rate over time with episodic bursts of mutations. The initiating factors for the bursts are unclear, although retrotransposon mobilization may contribute. The examined cell lines constitute a resource of live experimental models of mutational processes, which potentially retain patterns of activity and regulation operative in primary human cancers.
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http://dx.doi.org/10.1016/j.cell.2019.02.012DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6424819PMC
March 2019

Identification of a quadruple mutation that confers tenofovir resistance in chronic hepatitis B patients.

Authors:
Eun-Sook Park Ah Ram Lee Doo Hyun Kim Jeong-Hoon Lee Jeong-Ju Yoo Sung Hyun Ahn Heewoo Sim Soree Park Hong Seok Kang Juhee Won Yea Na Ha Gu-Choul Shin So Young Kwon Yong Kwang Park Byeong-Sun Choi Yun Bin Lee Nakcheol Jeong Yohan An Young Seok Ju Su Jong Yu Hee Bok Chae Kyung-Sang Yu Yoon Jun Kim Jung-Hwan Yoon Fabien Zoulim Kyun-Hwan Kim

J Hepatol 2019 06 20;70(6):1093-1102. Epub 2019 Feb 20.

Department of Pharmacology and Center for Cancer Research and Diagnostic Medicine, IBST, School of Medicine, Konkuk University, Seoul, Republic of Korea; KU Open Innovation Center, Konkuk University, Seoul, Republic of Korea; Research Institute of Medical Sciences, Konkuk University, Seoul, Republic of Korea. Electronic address:

Background & Aims: Tenofovir disoproxil fumarate (TDF) is one the most potent nucleot(s)ide analogues for treating chronic hepatitis B virus (HBV) infection. Phenotypic resistance caused by genotypic resistance to TDF has not been reported. This study aimed to characterize HBV mutations that confer tenofovir resistance.

Methods: Two patients with viral breakthrough during treatment with TDF-containing regimens were prospectively enrolled. The gene encoding HBV reverse transcriptase was sequenced. Eleven HBV clones harboring a series of mutations in the reverse transcriptase gene were constructed by site-directed mutagenesis. Drug susceptibility of each clone was determined by Southern blot analysis and real-time PCR. The relative frequency of mutants was evaluated by ultra-deep sequencing and clonal analysis.

Results: Five mutations (rtS106C [C], rtH126Y [Y], rtD134E [E], rtM204I/V, and rtL269I [I]) were commonly found in viral isolates from 2 patients. The novel mutations C, Y, and E were associated with drug resistance. In assays for drug susceptibility, the IC value for wild-type HBV was 3.8 ± 0.6 µM, whereas the IC values for CYE and CYEI mutants were 14.1 ± 1.8 and 58.1 ± 0.9 µM, respectively. The IC value for wild-type HBV was 30 ± 0.5 µM, whereas the IC values for CYE and CYEI mutants were 185 ± 0.5 and 790 ± 0.2 µM, respectively. Both tenofovir-resistant mutants and wild-type HBV had similar susceptibility to the capsid assembly modulator NVR 3-778 (IC <0.4 µM vs. IC = 0.4 µM, respectively).

Conclusions: Our study reveals that the quadruple (CYEI) mutation increases the amount of tenofovir required to inhibit HBV by 15.3-fold in IC and 26.3-fold in IC. These results demonstrate that tenofovir-resistant HBV mutants can emerge, although the genetic barrier is high.

Lay Summary: Tenofovir is the most potent nucleotide analogue for the treatment of chronic hepatitis B virus infection and there has been no hepatitis B virus mutation that confers >10-fold resistance to tenofovir up to 8 years. Herein, we identified, for the first time, a quadruple mutation that conferred 15.3-fold (IC) and 26.3-fold (IC) resistance to tenofovir in 2 patients who experienced viral breakthrough during tenofovir treatment.
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http://dx.doi.org/10.1016/j.jhep.2019.02.006DOI Listing
June 2019
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