@article{LauGossenLendleinetal.2022,
  author    = {Lau, Skadi and Gossen, Manfred and Lendlein, Andreas and Jung, Friedrich},
  title     = {Differential sensitivity of assays for determining vein endothelial cell senescence},
  series = {Clinical hemorheology and microcirculation : blood flow and vessels},
  volume    = {81},
  journal   = {Clinical hemorheology and microcirculation : blood flow and vessels},
  number    = {3},
  publisher = {IOS Press},
  address   = {Amsterdam},
  issn      = {1386-0291},
  doi       = {10.3233/CH-211294},
  pages     = {191 -- 203},
  year      = {2022},
  abstract  = {In vivo endothelialization of polymer-based cardiovascular implant materials is a promising strategy to reduce the risk of platelet adherence and the subsequent thrombus formation and implant failure. However, endothelial cells from elderly patients are likely to exhibit a senescent phenotype that may counteract endothelialization. The senescence status of cells should therefore be investigated prior to implantation of devices designed to be integrated in the blood vessel wall. Here, human umbilical vein endothelial cells (HUVEC) were cultivated up to passage (P) 4, 10 and 26/27 to determine the population doubling time and the senescence status by four different methods. Determination of the senescence-associated beta-galactosidase activity (SA-beta-Gal) was carried out by colorimetric staining and microscopy (i), as well as by photometric quantification (ii), and the expression of senescence-associated nuclear proteins p16 and p21 as well as the proliferation marker Ki67 was assessed by immunostaining (iii), and by flow cytometry (iv). The population doubling time of P27-cells was remarkably greater (103 +/- 65 h) compared to P4-cells (24 +/- 3 h) and P10-cell (37 +/- 15 h). Among the four different methods tested, the photometric SA-beta-Gal activity assay and the flow cytometric determination of p16 and Ki67 were most effective in discriminating P27-cells from P4- and P10-cells. These methods combined with functional endothelial cell analyses might aid predictions on the performance of implant endothelialization in vivo.},
  language  = {en}
}
@article{MoradianGossenLendlein2022,
  author    = {Moradian, Hanieh and Gossen, Manfred and Lendlein, Andreas},
  title     = {Co-delivery of genes can be confounded by bicistronic vector design},
  series = {MRS Communications},
  volume    = {12},
  journal   = {MRS Communications},
  number    = {2},
  publisher = {Springer},
  address   = {Heidelberg},
  issn      = {2159-6859},
  doi       = {10.1557/s43579-021-00128-7},
  pages     = {145 -- 153},
  year      = {2022},
  abstract  = {Maximizing the efficiency of nanocarrier-mediated co-delivery of genes for co-expression in the same cell is critical for many applications. Strategies to maximize co-delivery of nucleic acids (NA) focused largely on carrier systems, with little attention towards payload composition itself. Here, we investigated the effects of different payload designs: co-delivery of two individual "monocistronic" NAs versus a single bicistronic NA comprising two genes separated by a 2A self-cleavage site. Unexpectedly, co-delivery via the monocistronic design resulted in a higher percentage of co-expressing cells, while predictive co-expression via the bicistronic design remained elusive. Our results will aid the application-dependent selection of the optimal methodology for co-delivery of genes.},
  language  = {en}
}
@article{MoradianRochAnthoferetal.2022,
  author    = {Moradian, Hanieh and Roch, Toralf and Anthofer, Larissa and Lendlein, Andreas and Gossen, Manfred},
  title     = {Chemical modification of uridine modulates mRNA-mediated proinflammatory and antiviral response in primary human macrophages},
  series = {Molecular therapy},
  volume    = {27},
  journal   = {Molecular therapy},
  publisher = {Cell Press},
  address   = {Cambridge},
  issn      = {2162-2531},
  doi       = {10.1016/j.omtn.2022.01.004},
  pages     = {854 -- 869},
  year      = {2022},
  abstract  = {In vitro transcribed (IVT)-mRNA has been accepted as a promising therapeutic modality. Advances in facile and rapid production technologies make IVT-mRNA an appealing alternative to protein- or virus-based medicines. Robust expression levels, lack of genotoxicity, and their manageable immunogenicity benefit its clinical applicability. We postulated that innate immune responses of therapeutically relevant human cells can be tailored or abrogated by combinations of 5'-end and internal IVT-mRNA modifications. Using primary human macrophages as targets, our data show the particular importance of uridine modifications for IVT-mRNA performance. Among five nucleotide modification schemes tested, 5-methoxy-uridine outperformed other modifications up to 4-fold increased transgene expression, triggering moderate proinflammatory and non-detectable antiviral responses. Macrophage responses against IVT-mRNAs exhibiting high immunogenicity (e.g., pseudouridine) could be minimized upon HPLC purification. Conversely, 5'-end modifications had only modest effects on mRNA expression and immune responses. Our results revealed how the uptake of chemically modified IVT-mRNA impacts human macrophages, responding with distinct patterns of innate immune responses concomitant with increased transient transgene expression. We anticipate our findings are instrumental to predictively address specific cell responses required for a wide range of therapeutic applications from eliciting controlled immunogenicity in mRNA vaccines to, e.g., completely abrogating cell activation in protein replacement therapies.},
  language  = {en}
}