Effective Drug Concentration and Selectivity Depends on Fraction of Primitive Cells

Lica, Jan; Wieczór, Miłosz; Grabe, Grzegorz; Jancz, Marta; Misiak, Majus; Gucwa, Katarzyna; Brankiewicz, Wioletta; Maciejewska, Natalia; Stupak, Anna; Bagiński, Maciej; Hellmann, Andrzej; Składanowski, Andrzej

doi:10.3390/ijms22094931

Cited by 36 publications

(37 citation statements)

References 87 publications

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“…3A). This is a documented phenomenon in cancer studies whereby a direct correlation has been drawn between starting cell densities and drug efficacy (31). In other works, TAT has documented measurable cytotoxicity above 10 µM (32) and our metabolic assays employed herein showed no to low cytotoxicity from TAT-CaM treatment only.…”

Section: Discussionsupporting

confidence: 68%

Utilization of a cell-penetrating peptide-adaptor for delivery of human papillomavirus protein E2 into cervical cancer cells to arrest cell growth and promote cell death

LeCher

Didier

Dickson

et al. 2022

Preprint

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Background: Human papillomavirus (HPV) is the causative agent of nearly all forms of cervical cancer, which can arise upon viral integration into the host genome and concurrent loss of viral regulatory gene E2. Gene-based delivery approaches show that E2 reintroduction reduces proliferative capacity and promotes apoptosis in vitro. This work explored if our calcium-dependent protein-based delivery system, TAT-CaM could deliver functional E2 protein directly into cervical cancer cells to limit proliferative capacity and induce cell death. Methods: TAT-CaM and the HPV16 E2 protein containing a CaM-binding sequence (CBS-E2) were expressed and purified from E. coli. Calcium-dependent binding kinetics were verified by Biolayer Interferometry. Equimolar TaT-CaM:CBS-E2 constructs were delivered into the HPV16+ SiHa cell line and uptake verified by confocal microscopy. Proliferative capacity was measured by MTS assay and cell death was measured by release of lactate dehydrogenase. As a control for specificity to HPV+ cells, human microvascular cells (HMECs) were used. Results: TAT-CaM bound CBS-E2 with high affinity in the presence of calcium and rapidly disassociated in its absence. After introduction by TAT-CaM, E2 was detected in cellular interiors by orthogonal projects taken at the depth of the nucleus. In dividing cells, E2 relocalized to regions associated with the mitotic spindle. Cells receiving a single daily dose of CBS-E2 for 4 days showed a significant reduction in metabolic activity at low doses and cell death at high doses compared to controls. This phenotype was retained for 7 days with no further treatments. When subcultured at day 12, treated cells regained their proliferative capacity. Conclusions: Using the TAT-CaM platform, bioactive E2 protein was delivered into living cervical cancer cells, inducing senescence and cell death in a time- and dose-dependent manner. These results suggest that this nucleic acid and virus-free delivery method could be harnessed to develop novel, effective protein therapeutics.

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Section: Discussionsupporting

confidence: 68%

Utilization of a cell-penetrating peptide-adaptor for delivery of human papillomavirus protein E2 into cervical cancer cells to arrest cell growth and promote cell death

LeCher

Didier

Dickson

et al. 2022

Preprint

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“…Nonetheless, the equal values for N/C ratios in both our cell populations were relatively low; thus, together with the findings from their qualitative ultrastructural observations, it can be suggested that the progenitor cells exhibited the features of an advanced differentiation stage. This assumption can be supported by the findings of Lica et al (2021) who characterized three sublines of A-549 cell line based on many aspects considering also the N/C ratio as follows: Primitive (N/C: 0.9–0.6), Standard (N/C: 0.6–0.2), and Mature (N/C: 0.2–0.05).…”

Section: Discussionmentioning

confidence: 53%

Rabbit Endothelial Progenitor Cells Derived From Peripheral Blood and Bone Marrow: An Ultrastructural Comparative Study

et al. 2022

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The present study was designed to compare the ultrastructure of early endothelial progenitor cells (EPCs) derived from rabbit peripheral blood (PB-EPCs) and bone marrow (BM-EPCs). After the cells had been isolated and cultivated up to passage 3, microphotographs obtained from transmission electron microscope were evaluated from qualitative and quantitative (unbiased stereological approaches) points of view. Our results revealed that both cell populations displayed almost identical ultrastructural characteristics represented by abundant cellular organelles dispersed in the cytoplasm. Moreover, the presence of very occasionally occurring mature endothelial-specific Weibel–Palade bodies (WPBs) confirmed their endothelial lineage origin. The more advanced stage of their differentiation was also demonstrated by the relatively low nucleus/cytoplasm (N/C) ratios (0.41 ± 0.19 in PB-EPCs; 0.37 ± 0.25 in BM-EPCs). Between PB-EPCs and BM-EPCs, no differences in proportions of cells occupied by nucleus (28.13 ± 8.97 versus 25.10 ± 11.48%), mitochondria (3.71 ± 1.33 versus 4.23 ± 1.00%), and lipid droplets (0.65 ± 1.01 versus 0.36 ± 0.40%), as well as in estimations of the organelles surface densities were found. The data provide the first quantitative evaluation of the organelles of interest in PB-EPCs and BM-EPCs, and they can serve as a research framework for understanding cellular function.

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“…Selectivity index is generated as a ratio of the lethal dose to the therapeutic dose of a given substance. The fibroblast and red blood cell IC 50 values represent the lethal dose (LD 50 ), while the IC 50 of a given drug is defined as the effective dose (ED 50 ) . An SI greater than or equal to 3 has been used as a criterion warranting continued studies of the candidate anticancer drug .…”

Section: Resultsmentioning

confidence: 99%

Molecularly Engineered Surfactin Analogues Induce Nonapoptotic-Like Cell Death and Increased Selectivity in Multiple Breast Cancer Cell Types

et al. 2023

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Surfactin, a negatively charged amphiphilic lipopeptide biosurfactant, is synthesized by the bacterium Bacillus subtilis. It consists of a cyclic heptapeptide and an 11−15C β-hydroxy fatty acid. To probe how the modification of the molecular skeleton of surfactin influences its selectivity and activity against breast cancer, six synthetic surfactins were generated. Modifications were accomplished by conjugating amine-functionalized molecules to the Glu and Asp carboxyl moieties of the heptapeptide. The resulting synthetic surfactins provided a diverse series of molecules with differences in charge, size, and hydrophilicity. After purification and structural analysis, insights into biological activity and specificity were generated for each compound. Dosedependent growth inhibition was determined for four tumorigenic breast cancer cell lines in monolayer and spheroid morphologies, as well as nontumorigenic fibroblasts and sheep erythrocytes, which were utilized to determine selectivity indices. Results indicated that two compounds, which have amplified anionic charge, had increased activity on breast cancer, with reduced activity on nontumorigenic fibroblasts and erythrocytes. Cationic derivative surf-ethylenediamine has increased activity on all cell lines tested. Novel correlations between dose−response activities and physicochemical properties of all compounds determined that there is a significant correlation between the critical micelle concentration and activity against multiple cell lines.

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Effective Drug Concentration and Selectivity Depends on Fraction of Primitive Cells

Cited by 36 publications

References 87 publications

Utilization of a cell-penetrating peptide-adaptor for delivery of human papillomavirus protein E2 into cervical cancer cells to arrest cell growth and promote cell death

Utilization of a cell-penetrating peptide-adaptor for delivery of human papillomavirus protein E2 into cervical cancer cells to arrest cell growth and promote cell death

Rabbit Endothelial Progenitor Cells Derived From Peripheral Blood and Bone Marrow: An Ultrastructural Comparative Study

Molecularly Engineered Surfactin Analogues Induce Nonapoptotic-Like Cell Death and Increased Selectivity in Multiple Breast Cancer Cell Types

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