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Tumor antigen and MHC expression in glioma cells for immunotherapeutic interventions

World Journal of Immunology 2013年第3期3卷 62-67页

作者： christina susanne mullins Alexander Walter Michael Schmitt Carl-Friedrich Classen Michael Linnebacher Pediatrics Department University Hospital Rostock 18057 Rostock Germany Department of General Surgery Molecular Oncology and Immunotherapy University Hospital Rostock 18057 Rostock Germany Department Internal Medicine V University Hospital Heidelberg 69120 Heidelberg Germany

AIM: To investigate the expression of tumor-antigens and major histocompatibility complex(MHC)-machinery components in glioblastoma multiforme cell lines flow cytometry staining methods were ***: Ten GBM cell lines(three commercially available: U-87 MG, U-138-MG and GMS-10 as well as seven newly established cell lines from individual patients in low-passages: HROG02, HROG04, HROG05, HROG06, HROG10, HROG13 and HROG17) were analyzed for expression of(Ⅰ) general and(Ⅱ) GBMrelated tumor antigens as well as of(Ⅲ) components of the MHC machinery by flow ***: All cell lines expressed MHC class?Ⅰ?with seven out of the ten being HLA-A02 positive. Four of the seven primary cell lines additionally expressedMHC class Ⅱ in a constitutive manner. Of note, after interferon gamma(IFN-γ) treatment, all seven cell lines expressed MHC class Ⅱ. The tumor associated antigens(TAA) EGFR and survivin were expressed at high levels in all cell lines; whereas MART-1, RHAMM, WT-1 and IL-13Rα were expressed by at least half of the cell lines and HER2/neu, MAGE-1 and tyrosinase were expressed only by few cell lines. However, all cell lines expressed at least two of the candidate antigens included into this ***: No obvious differences between commercially available and newly-established cell lines were observed. Thus, the latter in low-passages are interesting for(therapy-) screening and immunotherapeutic strategies.

关键词： Tumor antigens Glioblastoma multiforme Major histocompatibility complex molecules Tumor models Cell lines

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Colorectal cancer vaccines: Tumor-associated antigens vs neoantigens

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World Journal of Gastroenterology 2018年第48期24卷 5418-5432页

作者： Sandra Wagner christina S mullins Michael Linnebacher Section of Molecular Oncology and Immunotherapy General SurgeryUniversity Medical Center

Therapeutic options for the treatment of colorectal cancer(CRC) are diverse but still not always satisfying. Recent success of immune checkpoint inhibition treatment for the subgroup of CRC patients suffering from hypermutated tumors suggests a permanent role of immune therapy in the clinical management of CRC. Substantial improvement in treatment outcome could be achieved by development of efficient patient-individual CRC vaccination strategies. This mini-review summarizes the current knowledge on the two general classes of targets: tumor-associated antigens(TAAs) and tumorspecific antigens. TAAs like carcinoembryonic antigen and melanoma associated antigen are present in and shared by a subgroup of patients and a variety of clinical studies examined the efficacy of different TAA-derived peptide vaccines. Combinations of several TAAs as the next step and the development of personalized TAA-based peptide vaccines are discussed. Improvements of peptidebased vaccines achievable by adjuvants and immunestimulatory chemotherapeutics are highlighted. Finally, we sum up clinical studies using tumor-specific antigens-in CRC almost exclusively neoantigens-which revealed promising results; particularly no severe adverse events were reported so far. Critical progress for clinical outcomes can be expected by individualizing neoantigen-based peptide vaccines and combining them with immunestimulatory chemotherapeutics and immune checkpoint inhibitors. In light of these data and latest developments, truly personalized neoantigen-based peptide vaccines can be expected to fulfill modern precision medicine's requirements and will manifest as treatment pillar for routine clinical management of CRC.

关键词： Cancer vaccines Colorectal neoplasm Immunotherapy Neoplasm antigen tumor-associated antigens tumor-specific antigens Neoantigen(s)

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Mouse models of colorectal cancer: Past, present and future perspectives

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World Journal of Gastroenterology 2020年第13期26卷 1394-1426页

作者： Florian Bürtin christina S mullins Michael Linnebacher Department of General VisceralVascular and Transplantation SurgeryUniversity Medical Center RostockUniversity of RostockRostock 18057Germany Department of Thoracic Surgery University Medical Center RostockUniversity of RostockRostock 18057Germany Molecular Oncology and Immunotherapy Department of GeneralVisceralVascular and Transplantation SurgeryUniversity Medical Center RostockRostock 18057Germany

Colorectal cancer(CRC)is the third most common diagnosed malignancy among both sexes in the United States as well as in the European *** the incidence and mortality rates in western,high developed countries are declining,reflecting the success of screening programs and improved treatment regimen,a rise of the overall global CRC burden can be observed due to lifestyle changes paralleling an increasing human development *** a growing insight into the biology of CRC and many therapeutic improvements in the recent decades,preclinical in vivo models are still indispensable for the development of new treatment *** the development of carcinogen-induced rodent models for CRC more than 80 years ago,a plethora of animal models has been established to study colon cancer *** tenuous invasiveness and metastatic behavior,these models are useful for chemoprevention studies and to evaluate colitis-related *** engineered mouse models(GEMM)mirror the pathogenesis of sporadic as well as inherited CRC depending on the specific molecular pathways activated or *** the vast majority of CRC GEMM lack invasiveness,metastasis and tumor heterogeneity,they still have proven useful for examination of the tumor microenvironment as well as systemic immune responses;thus,supporting development of new therapeutic *** of metastatic disease by orthotopic injection of CRC cell lines is possible,but the so generated models lack genetic diversity and the number of suited cell lines is very ***-derived xenografts,in contrast,maintain the pathological and molecular characteristics of the individual patient's CRC after subcutaneous implantation into immunodeficient mice and are therefore most reliable for preclinical drug development–even in comparison to GEMM or cell line-based ***,subcutaneous patient-derived xenograft models are less suitable for studying most aspects of the tumor micr

关键词： Colorectal cancer Mouse models Patient-derived xenografts Carcinogen-induced models Genetically engineered mouse models Preclinical drug development

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A global assessment of recent trends in gastrointestinal cancer and lifestyle-associated risk factors

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Cancer Communications 2021年第11期41卷 1137-1151页

作者： Lili Lu christina S.mullins Clemens Schafmayer Sebastian Zeißig Michael Linnebacher Department of General Surgery Molecular Oncology and ImmunotherapyRostock University Medical CenterRostock 18057Germany Department of Medicine I University Hospital Carl Gustav CarusTechnische Universität(TU)DresdenDresden 01307Germany

Background:Gastrointestinal(GI)cancers were responsible for 26.3%of cancer cases and 35.4%of deaths worldwide in *** study aimed to analyze the global incidence,mortality,prevalence,and contributing risk factors of the 6 major GI cancer entities[esophageal cancer(EC),gastric cancer(GC),liver cancer(LC),pancreatic cancer(PC),colon cancer,and rectal cancer].Methods:Using the Global Cancer Observatory and the Global Health Observatory databases,we reviewed the current GI cancer incidence,prevalence,and mortality,analyzed the association of GI cancer prevalence with national human development indices(HDIs),identified the contributing risk factors,and estimated developing age-and sex-specific trends in incidence and ***:In 2020,the trend in age-standardized rate of incidence of GI cancers closely mirrored that of mortality,with the highest rates of LC,EC,and GC in Asia and of colorectal cancer(CRC)and PC mainly in *** and mortality were positively,but the mortality-to-incidence ratio(MIR)was inversely correlated with the national HDI *** MIRs in developing countries likely reflected the lack of preventive strategies and effective *** cancer prevalence was highest in Europe and was also positively correlated with HDIs and lifestyle-associated risk factors,such as alcohol consumption,smoking,obesity,insufficient physical activity,and high blood cholesterol level,but negatively correlated with hypertension and *** of EC were consistently and those of GC mostly decreasing,whereas incidences of CRC were increasing in most countries/regions,especially in the younger *** of LC and PC were also increasing in all age-gender populations except for younger *** were decreasing for EC,GC,and CRC in most countries/regions, and age-specific trends were observed in PC and LC with adecrease in the younger but an increase in the older ***: On the global scale, higher GI c

关键词： annual percentage change(APC) diabetes gastrointestinal(GI)cancer high blood cholesterol human development indices(HDI) hypertension incidence lifestyle mortality prevalence

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Recent advances in diagnosis and treatment of gastroenteropancreatic neuroendocrine neoplasms

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World Journal of Gastrointestinal Surgery 2022年第5期14卷 383-396页

作者： Meng Dai christina S mullins Lili Lu Guido Alsfasser Michael Linnebacher Clinic of General Surgery Molecular Oncology and ImmunotherapyRostock University Medical Center18057 RostockGermany Clinic of General Surgery Rostock University Medical Center18057 RostockGermany

Gastroenteropancreatic neuroendocrine neoplasms(GEP-NENs)are a rare group of tumors originating from neuroendocrine cells of the digestive *** incidence has increased over the last *** specific pathogenetic mechanisms underlying GEP-NEN development have not been completely *** GEP-NENs are usually asymptomatic;some grow slowly and thus impede early diagnosis,which ultimately results in a high rate of ***,many GEP-NEN patients present with later staged *** hereby,research attention for diagnosis and treatment for GEP-NENs increased in recent *** result of which is great progress in clinical diagnosis and *** to the most recent clinical guidelines,improved grading standards can accurately define poorly differentiated grade 3 neuroendocrine tumors and neuroendocrine carcinomas(NECs),which are subclassified into large and small cell *** different functional imaging methods facilitates precise *** expression of somatostatin receptors helps to predict *** analyses of mutations affecting death domain associated protein(DAXX),multiple endocrine neoplasia type 1(MEN 1),alpha thalassemia/intellectual disability syndrome X-linked(ATRX),retinoblastoma transcriptional corepressor 1(RB 1),and mothers against decapentaplegic homolog 4(SMAD 4)help distinguishing grade 3 NENs from poorly differentiated *** aim of this review is to summarize the latest research progress on diagnosis and treatment of GEP-NENs.

关键词： GEP-NENs Functional imaging Peptide receptor radionuclide therapy Targeting agents Immune checkpoint inhibitors Genetic mutations

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Human endogenous retroviruses and cancer:Causality and therapeutic possibilities

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World Journal of Gastroenterology 2012年第42期18卷 6027-6035页

作者： christina S mullins Michael Linnebacher Department of General ThoracicVascular and Transplantation SurgerySection Molecular Oncology and ImmunotherapyUniversity of RostockSchillingallee 3518057 RostockGermany

A substantial part of the human genome is derived from transposable elements;remnants of ancient retroviral *** estimates set the percentage of human endogenous retroviruses(HERVs) in the genome at 8%.For the most part,the interplay between mutations,epigenetic mechanisms and posttranscriptional regulations silence HERVs in somatic *** first highlight mechanisms by which activation of members of several HERV families may be associated with tumor development before discussing the arising chances for both diagnosis and *** has been shown that at least in some cases,tumor cells expressing HERV open reading frames(ORFs) thus gain tumor-promoting ***,since these proteins are not expressed in healthy tissues,they become prime target *** potential pharmacological interest are the prevention of HERV transposition,the inhibition of HERV-encoded protein expression and the interference with these proteins' *** from recent studies unequivocally proves that HERV ORFs represent a very interesting source of novel tumor-specific antigens with even the potential to surpass entity *** development of new tumor(immune-) therapies is a very active field and true tumor-specific targets are of outstanding interest since they minimize the risk of autoimmunity and could reduce side ***,we postulate on main future research streams in order to stimulate discussion on this hot topic.

关键词： Human endogenous retroviruses Gastro-intestinal cancer Therapeutic targets Tumor-specificantigens Tumorigenesis

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Establishment,functional and genetic characterization of a colon derived large cell neuroendocrine carcinoma cell line

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World Journal of Gastroenterology 2018年第33期24卷 3749-3759页

作者： Michael Gock christina S mullins Christine Harnack Friedrich Prall Robert Ramer Anja Goder Oliver H Kramer Ernst Klar Michael Linnebacher Department of General Surgery University of RostockRostock 18055Germany Department of General Surgery Section of Molecular Oncology and ImmunotherapyUniversity of RostockRostock 18055Germany Institute of Pathology University of RostockRostock 18055Germany Institute of Pharmacology University of RostockRostock 18055Germany Institute of Toxicology University Medical Center MainzMainz 55131Germany

AIM To establish cell line and patient-derived xenograft(PDX) models for neuroendocrine carcinomas(NEC) which is highly desirable for gaining insight into tumor development as well as preclinical research includingbiomarker testing and drug response *** Cell line establishment was conducted from direct in vitro culturing of colonic NEC tissue(HROC57). A PDX could also successfully be established from vitally frozen tumor samples. Morphological features, invasive and migratory behavior of the HROC57 cells as well as expression of neuroendocrine markers were vastly analyzed. Phenotypic analysis was done by microscopy and multicolor flow cytometry. The extensive molecular-pathological profiling included mutation analysis, assessment of chromosomal and microsatellite instability; and in addition, fingerprinting(i.e., STR analysis) was performed from the cell line in direct comparison to primary patient-derived tissues and the PDX model established. Drug responsiveness was examined for a panel of chemotherapeutics in clinical use for the treatment of solid *** The established cell line HROC57 showed distinct morphological and molecular features of a poorly differentiated large-cell NEC with KI-67 > 50%. Molecular-pathological analysis revealed a Cp G island promoter methylation positive cell line with microsatellite instability being absent. The following mutation profile was observed: KRAS(wt), BRAF(mut). A high sensitivity to etoposide, cisplatin and 5-FU could be demonstrated while it was more resistant towards rapamycin. CONCLUSION We successfully established and characterized a novel patient-derived NEC cell line in parallel to a PDX model as a useful tool for further analysis of the biological characteristics and for development of novel diagnostic and therapeutic options for NEC.

关键词： Patient-derived tumor model Large cell neuroendocrine carcinoma Individualized medicine

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Establishment,functional and genetic characterization of three novel patient-derived rectal cancer cell lines

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World Journal of Gastroenterology 2018年第43期24卷 4880-4892页

作者： Michael Gock christina S mullins Carina Bergner Friedrich Prall Robert Ramer Anja Goder Oliver H Kramer Falko Lange Bernd J Krause Ernst Klar Michael Linnebacher Department of General Surgery University Medical CenterRostock 18055Germany Section of Molecular Oncology and Immunotherapy University MedicalCenterRostock 18055Germany Department of NuclearMedicine University Medical CenterRostock 18055Germany Institute of Pathology University MedicalCenterRostock 18055Germany Institute of Pharmacology University Medical CenterRostock 18055Germany Institute of Toxicology University Medical Center MainzMainz 55131Germany Oscar-Langendorff-Institute of Physiology University Medical CenterRostock 18055Germany

AIM To establish patient-individual tumor models of rectal cancer for analyses of novel biomarkers, individual response prediction and individual therapy *** Establishment of cell lines was conducted by direct in vitro culturing and in vivo xenografting with subsequent in vitro culturing. Cell lines were in-depth characterized concerning morphological features, invasive and migratory behavior, phenotype, molecular profile including mutational analysis, protein expression, and confirmation of origin by DNA fingerprint. Assessment of chemosensitivity towards an extensive range of current chemotherapeutic drugs and of radiosensitivity was performed including analysis of a combined radioand chemotherapeutic treatment. In addition, glucose metabolism was assessed with 18 F-fluorodeoxyglucose(FDG) and proliferation with 18 *** We describe the establishment of ultra-low passage rectal cancer cell lines of three patients suffering from rectal cancer. Two cell lines(HROC126, HROC284 Met) were established directly from tumor specimens while HROC239 T0 M1 was established subsequent to xenografting of the tumor. Molecular analysis classified all three cell lines as CIMP-0/non-MSI-H(sporadic standard) type. Mutational analysis revealed following mutational profiles: HROC126: APC^(wt), TP53^(wt), KRAS^(wt), BRAF^(wt), PTEN^(wt); HROC239 T0 M1: APC^(mut), P53^(wt), KRAS^(mut), BRAF^(wt), PTEN^(mut) and HROC284 Met: APC^(wt), P53^(mut), KRAS^(mut), BRAF^(wt), PTEN^(mut). All cell lines could be characterized as epithelial(EpCAM+) tumor cells with equivalent morphologic features and comparable growth kinetics. The cell lines displayed a heterogeneous response toward chemotherapy, radiotherapy and their combined application. HROC126 showed a highly radio-resistant phenotype and HROC284 Met was more susceptible to a combined radiochemotherapy than HROC126 and HROC239 T0 M1. Analysis of 18 F-FDG uptake displayed a markedly reduced FDG uptake of all th

关键词： Patient-derived tumor model Rectal cancer ^(18)F-fluorodeoxyglucose ^(18)F-fluorothymidine FOLFOX FOLFIRI Personalized medicine

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Integrating Culture-based Antibiotic Resistance Profiles with Whole-genome Sequencing Data for 11,087 Clinical Isolates

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Genomics, Proteomics & Bioinformatics 2019年第2期17卷 169-182页

作者： Valentina Galata Cédric C. Laczny christina Backes Georg Hemmrich-Stanisak susanne Schmolke Andre Franke Eckart Meese Mathias Herrmann Lutz von Müller Achim Plum Rolf Mü ller Cord Stahler Andreas E. Posch Andreas Keller Chair for Clinical Bioinformatics Saarland University 66123 Saarbrücken Germany Helmholtz Center for Infection Research and Pharmaceutical Biotechnology (HZI) Saarland University 66123 Saarbrücken Germany Institute of Clinical Molecular Biology Christian-Albrechts University of Kiel 24105 Kiel Germany Siemens Healthcare GmbH Strategy and Innovation 91052 Erlangen Germany Department of Human Genetics Saarland University 66421 Homburg Germany Institute of Medical Microbiology and Hygiene Saarland University 66421 Homburg Germany Ares Genetics GmbH 1030 Vienna Austria Curetis GmbH 71088 Holzgerlingen Germany Department of Pharmacy Pharmaceutical Biotechnology Saarland University 66123 Saarbrücken Germany Department of Microbial Natural Products Helmholtz-Institute for Pharmaceutical Research Saarland (HIPS) Saarland University 66123 Saarbrücken Germany

Emerging antibiotic resistance is a major global health threat. The analysis of nucleic acid sequences linked to susceptibility phenotypes facilitates the study of genetic antibiotic resistance determinants to inform molecular diagnostics and drug development. We collected genetic data (11,087 newly-sequenced whole genomes) and culture-based resistance profiles (10,991 out of the 11,087 isolates comprehensively tested against 22 antibiotics in total) of clinical isolates including 18 main species spanning a time period of 30 years. Species and drug specific resistance patterns were observed including increased resistance rates for Acinetobacter baumannii to carbapenems and for Escherichia coli to fluoroquinolones. Species-level pan-genomes were constructed to reflect the genetic repertoire of the respective species,including conserved essential genes and known resis-tance factors. Integrating phenotypes and genotypes through species-level pan-genomes allowed to infer gene–drug resistance associations using statistical testing. The isolate collection and the analysis results have been integrated into GEAR-base,a resource available for academic research use free of charge at https://***.

关键词： Antibiotic resistance Whole-genome sequencing Bacteria Pan-genome

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Biocompatibility and Antibacterial Effects of 6-Deoxy-6-Aminoethyleneamino Cellulose

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Journal of Biosciences and Medicines 2018年第1期6卷 51-62页

作者： susanne Finger Michael Zieger Cornelia Wiegand Tim Liebert Thomas Heinze Peter Elsner Uta-christina Hipler Department of Dermatology University Medical Center Jena Germany Center of Excellence for Polysaccharide Research Institute of Organic Chemistry and Macromolecular Chemistry Friedrich Schiller University Jena Germany

Recently, there is a need of alternatives to antibiotics due to increasing antibiotic-resistant microorganism. Promising classes of bioactive polymers are 6-deoxy-6-amino cellulose derivatives. The purpose of the study was the assessment of the biocompatibility of 6-deoxy-6-aminoethyleneamino cellulose (AEAC) with different degree of substitution (DS). HaCaT keratinocyte cell viability was analyzed by measuring the cellular ATP content. The antibacterial activity against Staphylococcus aureus and Klebsiella pneumoniae was examined by microplate laser nephelometry. Thus, the ratio of half-maximal lethal concentration (LC50) and half-maximal inhibitory concentration (IC50) was calculated and described as biocompatibility index. The study revealed that biocompatibility of AEAC depends on the DS. AEAC of low DS (0.3) showed the best biocompatibility.

关键词： 6-Deoxy-6-Aminoethyleneamino Celluloses Antibacterial Bacteria Biocompatibility Biomacromolecule Keratinocyte

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