Serum from calorie-restricted animals delays senescence and extends the lifespan of normal human fibroblasts in vitro

The COVID-19 pandemic is still active around the globe despite the newly introduced vaccines. Hence, finding effective medications or repurposing available ones could offer great help during this serious situation. During our anti-COVID-19 investigation of microbial natural products (MNPs), we came across α-rubromycin, an antibiotic derived from Streptomyces collinus ATCC19743, which was able to suppress the catalytic activity (IC50 = 5.4 µM and Ki = 3.22 µM) of one of the viral key enzymes (i.e., MPro). However, it showed high cytotoxicity toward normal human fibroblasts (CC50 = 16.7 µM). To reduce the cytotoxicity of this microbial metabolite, we utilized a number of in silico tools (ensemble docking, molecular dynamics simulation, binding free energy calculation) to propose a novel scaffold having the main pharmacophoric features to inhibit MPro with better drug-like properties and reduced/minimal toxicity. Nevertheless, reaching this novel scaffold synthetically is a time-consuming process, particularly at this critical time. Instead, this scaffold was used as a template to explore similar molecules among the FDA-approved medications that share its main pharmacophoric features with the aid of pharmacophore-based virtual screening software. As a result, cromoglicic acid (aka cromolyn) was found to be the best hit, which, upon in vitro MPro testing, was 4.5 times more potent (IC50 = 1.1 µM and Ki = 0.68 µM) than α-rubromycin, with minimal cytotoxicity toward normal human fibroblasts (CC50 > 100 µM). This report highlights the potential of MNPs in providing unprecedented scaffolds with a wide range of therapeutic efficacy. It also revealed the importance of cheminformatics tools in speeding up the drug discovery process, which is extremely important in such a critical situation.

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Design, Synthesis, and Cytotoxicity Assessment of [64Cu]Cu-NOTA-Terpyridine Platinum Conjugate: A Novel Chemoradiotherapeutic Agent with Flexible Linker

Nanomaterials ◽

10.3390/nano11092154 ◽

2021 ◽

Vol 11 (9) ◽

pp. 2154

Author(s):

Meysam Khosravifarsani ◽

Samia Ait-Mohand ◽

Benoit Paquette ◽

Léon Sanche ◽

Brigitte Guérin

Keyword(s):

Colorectal Cancer ◽

Cancer Cells ◽

Human Fibroblasts ◽

Fold Increase ◽

Radiation Energy ◽

Design Synthesis ◽

Quadruplex Dna ◽

Low Energy Electrons ◽

Normal Human

Maximum benefits of chemoradiation therapy with platinum-based compounds are expected if the radiation and the drug are localized simultaneously in cancer cells. To optimize this concomitant effect, we developed the novel chemoradiotherapeutic agent [64Cu]Cu-NOTA-C3-TP by conjugating, via a short flexible alkyl chain spacer (C3), a terpyridine platinum (TP) moiety to a NOTA chelator complexed with copper-64 (64Cu). The decay of 64Cu produces numerous low-energy electrons, enabling the 64Cu-conjugate to deliver radiation energy close to TP, which intercalates into G-quadruplex DNA. Accordingly, the in vitro internalization kinetic and the cytotoxic activity of [64Cu]Cu-NOTA-C3-TP and its derivatives were investigated with colorectal cancer (HCT116) and normal human fibroblast (GM05757) cells. Radiolabeling by 64Cu results in a >55,000-fold increase of cytotoxic potential relative to [NatCu]Cu-NOTA-C3-TP at 72 h post administration, indicating a large additive effect between 64Cu and the TP drug. The internalization and nucleus accumulation of [64Cu]Cu-NOTA-C3-TP in the HCT116 cells were, respectively, 3.1 and 6.0 times higher than that for GM05757 normal human fibroblasts, which is supportive of the higher efficiency of the [64Cu]Cu-NOTA-C3-TP for HCT116 cancer cells. This work presents the first proof-of-concept study showing the potential use of the [64Cu]Cu-NOTA-C3-TP conjugate as a targeted chemoradiotherapeutic agent to treat colorectal cancer.

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Chromosome 7 suppresses indefinite division of nontumorigenic immortalized human fibroblast cell lines KMST-6 and SUSM-1

Molecular and Cellular Biology ◽

10.1128/mcb.13.10.6036-6043.1993 ◽

1993 ◽

Vol 13 (10) ◽

pp. 6036-6043

Author(s):

T Ogata ◽

D Ayusawa ◽

M Namba ◽

E Takahashi ◽

M Oshimura ◽

...

Keyword(s):

Cell Lines ◽

Human Cell ◽

Human Fibroblasts ◽

Fibroblast Cell ◽

Chromosome 7 ◽

In Vitro Mutagenesis ◽

Human Cell Lines ◽

First Case ◽

Normal Human

Using nontumorigenic immortalized human cell lines KMST-6 (KMST) and SUSM-1 (SUSM), we attempted to identify the chromosome that carries a putative senescence-related gene(s). These cell lines are the only ones that have been established independently from normal human diploid fibroblasts following in vitro mutagenesis. We first examined restriction fragment length polymorphisms on each chromosome of these immortalized cell lines and their parental cell lines and found specific chromosomal alterations common to these cell lines (a loss of heterozygosity in KMST and a deletion in SUSM) on the long arm of chromosome 7. In addition to these, we also found that introduction of chromosome 7 into these cell lines by means of microcell fusion resulted in the cessation of cell division, giving rise to cells resembling cells in senescence. Introduction of other chromosomes, such as chromosomes 1 and 11, on which losses of heterozygosity were also detected in one of the cell lines (KMST), to either KMST or SUSM cells or of chromosome 7 to several tumor-derived cell lines had no effect on their division potential. These results strongly suggest that a gene(s) affecting limited-division potential or senescence of normal human fibroblasts is located on chromosome 7, probably at the long arm of the chromosome, representing the first case in which a specific chromosome reverses the immortal phenotype of otherwise normal human cell lines.

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A Novel Tumor Suppressor, REIC/Dkk-3 Gene Identified by Our In Vitro Transformation Model of Normal Human Fibroblasts Works as a Potent Therapeutic Anti-tumor Agent

Advances in Experimental Medicine and Biology - Human Cell Transformation ◽

10.1007/978-1-4614-0254-1_17 ◽

2011 ◽

pp. 209-215 ◽

Cited By ~ 6

Author(s):

Masakiyo Sakaguchi ◽

Nam-ho Huh ◽

Masayoshi Namba

Keyword(s):

Tumor Suppressor ◽

Human Fibroblasts ◽

Transformation Model ◽

In Vitro Transformation ◽

Normal Human ◽

Tumor Agent

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Effects of Isologous and Nonisologous Nucleic Acids on the Ageing of Normal Human Fibroblasts in vitro

Pathology - Research and Practice ◽

10.1016/s0344-0338(78)80082-x ◽

1978 ◽

Vol 163 (2) ◽

pp. 137-147

Author(s):

J. Mellgren ◽

A.-M. Malmberg ◽

B. Rynell

Keyword(s):

Nucleic Acids ◽

Human Fibroblasts ◽

Normal Human

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Transformation of Normal Human Fibroblasts into Histologically Malignant Tissue in vitro

Science ◽

10.1126/science.123.3195.502-b ◽

1956 ◽

Vol 123 (3195) ◽

pp. 502-503

Author(s):

Joseph Leighton ◽

Ira Kline ◽

Henry C. Orr

Keyword(s):

Human Fibroblasts ◽

Malignant Tissue ◽

Normal Human

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High and low affinity binding sites for Concanavalin A on normal human fibroblasts in vitro

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(77)90959-7 ◽

1977 ◽

Vol 76 (4) ◽

pp. 1027-1035 ◽

Cited By ~ 14

Author(s):

Martha Feller ◽

Charles Richardson ◽

W. David Behnke ◽

Eric Gruenstein

Keyword(s):

Concanavalin A ◽

Binding Sites ◽

Human Fibroblasts ◽

Affinity Binding ◽

Normal Human

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Selective adhesion and impaired adhesive properties of transformed cells

Journal of Cell Science ◽

10.1242/jcs.33.1.121 ◽

1978 ◽

Vol 33 (1) ◽

pp. 121-132

Author(s):

M. Brugmans ◽

J.J. Cassiman ◽

H. van den Berghe

Keyword(s):

Cell Lines ◽

Animal Cell ◽

Human Fibroblasts ◽

Transformed Cells ◽

Cellular Interactions ◽

Human Tumour ◽

Adhesive Properties ◽

Normal Human ◽

Adhesive Interactions

Quantitative studies on the adhesive properties of transformed cells have yielded inconclusive and sometimes contradictory results. The present investigation has examined adhesive interactions between normal human fibroblasts, established as well as virus-transformed animal cell lines, and human tumour-derived cell lines by the cell-cell layer binding assay. The results of these investigations indicate that adhesive selectivity can be observed between normal human fibroblasts and 2 human tumour-derived cell lines, providing an in vitro system to study cell surface components involved in cellular interactions between normal and malignant cells. In addition it is demonstrated that cell layers of transformed cells form a poorly adhesive substratum for both trypsinized normal and transformed cells. Furthermore, it is confirmed that the adhesive properties of transformed cells, including adhesive selectivity, are affected by the dissociation procedure (trypsin or EDTA). In view of the observations made by other investigators, the present results suggest that transformed cells display adhesive properties which can be quantitatively and reproducibly measured but which are modulated by the dissociation procedure as well as by the configuration in which the cells are at the time of the assay.

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