@article{HausmannFalkNeitzeletal.2021, author = {Hausmann, Michael and Falk, Martin and Neitzel, Charlotte and Hofmann, Andreas and Biswas, Abin and Gier, Theresa and Falkova, Iva and Heermann, Dieter W. and Hildenbrand, Georg}, title = {Elucidation of the Clustered Nano-Architecture of Radiation-Induced DNA Damage Sites and Surrounding Chromatin in Cancer Cells: A Single Molecule Localization Microscopy Approach}, series = {International Journal of Molecular Sciences}, volume = {22}, journal = {International Journal of Molecular Sciences}, number = {7}, publisher = {MDPI AG}, issn = {1422-0067}, doi = {https://doi.org/10.3390/ijms22073636}, year = {2021}, abstract = {In cancer therapy, the application of (fractionated) harsh radiation treatment is state of the art for many types of tumors. However, ionizing radiation is a "double-edged sword"—it can kill the tumor but can also promote the selection of radioresistant tumor cell clones or even initiate carcinogenesis in the normal irradiated tissue. Individualized radiotherapy would reduce these risks and boost the treatment, but its development requires a deep understanding of DNA damage and repair processes and the corresponding control mechanisms. DNA double strand breaks (DSBs) and their repair play a critical role in the cellular response to radiation. In previous years, it has become apparent that, beyond genetic and epigenetic determinants, the structural aspects of damaged chromatin (i.e., not only of DSBs themselves but also of the whole damage-surrounding chromatin domains) form another layer of complex DSB regulation. In the present article, we summarize the application of super-resolution single molecule localization microscopy (SMLM) for investigations of these structural aspects with emphasis on the relationship between the nano-architecture of radiation-induced repair foci (IRIFs), represented here by γH2AX foci, and their chromatin environment. Using irradiated HeLa cell cultures as an example, we show repair-dependent rearrangements of damaged chromatin and analyze the architecture of γH2AX repair clusters according to topological similarities. Although HeLa cells are known to have highly aberrant genomes, the topological similarity of γH2AX was high, indicating a functional, presumptively genome type-independent relevance of structural aspects in DSB repair. Remarkably, nano-scaled chromatin rearrangements during repair depended both on the chromatin domain type and the treatment. Based on these results, we demonstrate how the nano-architecture and topology of IRIFs and chromatin can be determined, point to the methodological relevance of SMLM, and discuss the consequences of the observed phenomena for the DSB repair network regulation or, for instance, radiation treatment outcomes.}, subject = {Krebszelle}, language = {en} } @article{BobkovaDepesLeeetal.2018, author = {Bobkova, Elizaveta and Depes, Daniel and Lee, Jin-Ho and Jezkova, Lucie and Falkova, Iva and Pagacova, Eva and Kopecna, Olga and Zadneprianetc, Mariia and Bacikova, Alena and Kulikova, Elena and Smirnova, Elena and Bulanova, Tatiana and Boreyko, Alla and Krasavin, Evgeny and Wenz, Frederik and Bestvater, Felix and Hildenbrand, Georg and Hausmann, Michael and Falk, Martin}, title = {Recruitment of 53BP1 Proteins for DNA Repair and Persistence of Repair Clusters Differ for Cell Types as Detected by Single Molecule Localization Microscopy}, series = {International Journal of Molecular Sciences}, volume = {19}, journal = {International Journal of Molecular Sciences}, number = {12}, publisher = {MDPI AG}, issn = {1422-0067}, doi = {https://doi.org/10.3390/ijms19123713}, year = {2018}, abstract = {DNA double stranded breaks (DSBs) are the most serious type of lesions introduced into chromatin by ionizing radiation. During DSB repair, cells recruit different proteins to the damaged sites in a manner dependent on local chromatin structure, DSB location in the nucleus, and the repair pathway entered. 53BP1 is one of the important players participating in repair pathway decision of the cell. Although many molecular biology details have been investigated, the architecture of 53BP1 repair foci and its development during the post-irradiation time, especially the period of protein recruitment, remains to be elucidated. Super-resolution light microscopy is a powerful new tool to approach such studies in 3D-conserved cell nuclei. Recently, we demonstrated the applicability of single molecule localization microscopy (SMLM) as one of these highly resolving methods for analyses of dynamic repair protein distribution and repair focus internal nano-architecture in intact cell nuclei. In the present study, we focused our investigation on 53BP1 foci in differently radio-resistant cell types, moderately radio-resistant neonatal human dermal fibroblasts (NHDF) and highly radio-resistant U87 glioblastoma cells, exposed to high-LET 15N-ion radiation. At given time points up to 24 h post irradiation with doses of 1.3 Gy and 4.0 Gy, the coordinates and spatial distribution of fluorescently tagged 53BP1 molecules was quantitatively evaluated at the resolution of 10-20 nm. Clusters of these tags were determined as sub-units of repair foci according to SMLM parameters. The formation and relaxation of such clusters was studied. The higher dose generated sufficient numbers of DNA breaks to compare the post-irradiation dynamics of 53BP1 during DSB processing for the cell types studied. A perpendicular (90°) irradiation scheme was used with the 4.0 Gy dose to achieve better separation of a relatively high number of particle tracks typically crossing each nucleus. For analyses along ion-tracks, the dose was reduced to 1.3 Gy and applied in combination with a sharp angle irradiation (10° relative to the cell plane). The results reveal a higher ratio of 53BP1 proteins recruited into SMLM defined clusters in fibroblasts as compared to U87 cells. Moreover, the speed of foci and thus cluster formation and relaxation also differed for the cell types. In both NHDF and U87 cells, a certain number of the detected and functionally relevant clusters remained persistent even 24 h post irradiation; however, the number of these clusters again varied for the cell types. Altogether, our findings indicate that repair cluster formation as determined by SMLM and the relaxation (i.e., the remaining 53BP1 tags no longer fulfill the cluster definition) is cell type dependent and may be functionally explained and correlated to cell specific radio-sensitivity. The present study demonstrates that SMLM is a highly appropriate method for investigations of spatiotemporal protein organization in cell nuclei and how it influences the cell decision for a particular repair pathway at a given DSB site.}, subject = {Genom}, language = {en} } @article{HildenbrandMetzlerPilarczyketal.2018, author = {Hildenbrand, Georg and Metzler, Philipp and Pilarczyk, G{\"o}tz and Bobu, Vladimir and Kriz, Wilhelm and Hosser, Hiltraud and Fleckenstein, Jens and Krufczik, Matthias and Bestvater, Felix and Wenz, Frederik and Hausmann, Michael}, title = {Dose enhancement effects of gold nanoparticles specifically targeting RNA in breast cancer cells}, series = {PLOS ONE}, volume = {13}, journal = {PLOS ONE}, number = {1}, editor = {Baptista, Pedro V.}, publisher = {Public Library of Science (PLoS)}, issn = {1932-6203}, doi = {https://doi.org/10.1371/journal.pone.0190183}, year = {2018}, subject = {Krebs, Medizin}, language = {en} } @article{MoserHildenbrandMuelleretal.2016, author = {Moser, Felipe and Hildenbrand, Georg and M{\"u}ller, Patrick and Al Saroori, Alexander and Biswas, Abin and Bach, Margund and Wenz, Frederik and Cremer, Christoph and Burger, Nina and Veldwijk, Marlon R. and Hausmann, Michael}, title = {Cellular Uptake of Gold Nanoparticles and Their Behavior as Labels for Localization Microscopy}, series = {Biophysical Journal}, volume = {110}, journal = {Biophysical Journal}, number = {4}, publisher = {Elsevier BV}, issn = {0006-3495}, doi = {https://doi.org/10.1016/j.bpj.2016.01.004}, pages = {947 -- 953}, year = {2016}, subject = {Krebs, Medizin}, language = {en} } @article{StuhlmuellerSchwarzFinsterleFeyetal.2015, author = {Stuhlm{\"u}ller, Michael and Schwarz-Finsterle, Jutta and Fey, Evelyn and Lux, Johannes and Bach, Margund and Cremer, Christoph and Hinderhofer, Katrin and Hausmann, Michael and Hildenbrand, Georg}, title = {In situ optical sequencing and structure analysis of a trinucleotide repeat genome region by localization microscopy after specific COMBO-FISH nano-probing}, series = {Nanoscale}, volume = {7}, journal = {Nanoscale}, number = {42}, publisher = {Royal Society of Chemistry (RSC)}, issn = {2040-3364}, doi = {https://doi.org/10.1039/C5NR04141D}, pages = {17938 -- 17946}, year = {2015}, subject = {Genom}, language = {en} } @article{BosiekHausmannHildenbrand2016, author = {Bosiek, Katharina and Hausmann, Michael and Hildenbrand, Georg}, title = {Perspectives on Comets, Comet-like Asteroids, and Their Predisposition to Provide an Environment That Is Friendly to Life}, series = {Astrobiology}, volume = {16}, journal = {Astrobiology}, number = {4}, publisher = {Mary Ann Liebert Inc}, issn = {1531-1074}, doi = {https://doi.org/10.1089/ast.2015.1354}, pages = {311 -- 323}, year = {2016}, subject = {Komet}, language = {en} } @article{HausmannWinklerHildenbrandetal.2003, author = {Hausmann, Michael and Winkler, Ralph and Hildenbrand, Georg and Finsterle, Jutta and Weisel, Andrea and Rapp, Alexander and Schmitt, Eberhard and Janz, Siegfried and Cremer, Christoph}, title = {COMBO-FISH: specific labeling of nondenatured chromatin targets by computer-selected DNA oligonucleotide probe combinations}, series = {BioTechniques}, volume = {35}, journal = {BioTechniques}, number = {3}, publisher = {Informa UK Limited}, issn = {0736-6205}, doi = {https://doi.org/10.2144/03353rr03}, pages = {564 -- 577}, year = {2003}, subject = {Genom}, language = {en} } @incollection{FalkWolinskyVeldwijketal.2020, author = {Falk, Martin and Wolinsky, Michael and Veldwijk, Marlon R. and Hildenbrand, Georg and Hausmann, Michael}, title = {Gold nanoparticle enhanced radiosensitivity of cells: considerations and contradictions from model systems and basic investigations of cell damaging for radiation therapy}, series = {Nanoparticle Enhanced Radiation Therapy}, booktitle = {Nanoparticle Enhanced Radiation Therapy}, publisher = {IOP Publishing}, isbn = {9780750323963}, doi = {https://doi.org/10.1088/978-0-7503-2396-3ch10}, pages = {10 -- 1}, year = {2020}, subject = {Krebs, Medizin}, language = {en} } @misc{LeeBobkovaGieretal.2019, author = {Lee, Jin-Ho and Bobkova, Elizaveta and Gier, Theresa and Gote, Martin and Schmidt-kaler, Fanz and Brieger, Emily and Maus, Emanuel and Krufczik, Matthias and Chojowski, Robert and Korn, Friederike and Sarah, Schumann and Scherthan, Harry and Falkova, Iva and Falk, Martin and Hausmann, Michael and Hildenbrand, Georg}, title = {Mechanisms and Challenges for Understanding Radiation Induced Changes in Chromatin Nanoarchitecture}, series = {DRO 2018}, journal = {DRO 2018}, doi = {https://doi.org/10.13140/RG.2.2.31391.20647}, year = {2019}, abstract = {The three-dimensional architecture of genomes acts as an additional level of mode for fundamental biological processes such as DNA damage response. In this context, nanoprobing and super-resolution microscopy are powerful methods for structural analyses of genomic targets in native chromatin of single cells at resolutions of single antibodies, proteins, histones, short DNA stretches, etc. We used multi-color chromatin nanoprobing and single molecule localization microscopy of established DNA damage and chromatin markers in 3D-conserved nuclei of different cell types exposed to various types and doses of ionizing radiation. Similarly, effect of gold nanoparticles on extent and direction of cellular radiation response was assessed. Our studies revealed the nanoarchitecture of damage foci with respect to γH2AX, Mre11 or 53BP1 and their molecular rearrangements during repair processes. Nanoscopy of genomic Alu resulted in linear-quadratic dose-effects for low to higher dose ranges and in changes of H3K9me3 distribution around Alu clusters upon radiation exposure. Preliminary results show post-irradiation time dependent changes in Alu chromatin. Moreover, gold nanoparticles incorporated into cells seem to act by an interplay of radiation enhancement and chromatin remodeling leading to altered radiosensitivity. Our results contribute to the understanding of cellular radiation responses, thereby laying the basis for improved biological dosimetry and radiotherapies in future.}, subject = {Genom}, language = {en} } @article{NgwaBoatengKumaretal.2017, author = {Ngwa, Wilfred and Boateng, Francis and Kumar, Rajiv and Irvine, Darrell J. and Formenti, Silvia and Ngoma, Twalib and Herskind, Carsten and Veldwijk, Marlon R. and Hildenbrand, Georg and Hausmann, Michael and Wenz, Frederik and Hesser, J{\"u}rgen}, title = {Smart Radiation Therapy Biomaterials}, series = {International Journal of Radiation Oncology*Biology*Physics}, volume = {97}, journal = {International Journal of Radiation Oncology*Biology*Physics}, number = {3}, publisher = {Elsevier BV}, issn = {0360-3016}, doi = {https://doi.org/10.1016/j.ijrobp.2016.10.034}, pages = {624 -- 637}, year = {2017}, subject = {Krebs, Medizin}, language = {en} }