Cellular effects and delivery propensity of penetratin is influenced by conjugation to parathyroid hormone fragment 1-34 in synergy with pH

Research output: Contribution to journalJournal articlepeer-review

Standard

Cellular effects and delivery propensity of penetratin is influenced by conjugation to parathyroid hormone fragment 1-34 in synergy with pH. / Kristensen, Mie; Nielsen, Line Hagner; Zor, Kinga; Boisen, Anja; Christensen, Malene Vinther; Berthelsen, Jens; Nielsen, Hanne Mørck.

In: Bioconjugate Chemistry, Vol. 29, No. 2, 2018, p. 371-381.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Kristensen, M, Nielsen, LH, Zor, K, Boisen, A, Christensen, MV, Berthelsen, J & Nielsen, HM 2018, 'Cellular effects and delivery propensity of penetratin is influenced by conjugation to parathyroid hormone fragment 1-34 in synergy with pH', Bioconjugate Chemistry, vol. 29, no. 2, pp. 371-381. https://doi.org/10.1021/acs.bioconjchem.7b00687

APA

Kristensen, M., Nielsen, L. H., Zor, K., Boisen, A., Christensen, M. V., Berthelsen, J., & Nielsen, H. M. (2018). Cellular effects and delivery propensity of penetratin is influenced by conjugation to parathyroid hormone fragment 1-34 in synergy with pH. Bioconjugate Chemistry, 29(2), 371-381. https://doi.org/10.1021/acs.bioconjchem.7b00687

Vancouver

Kristensen M, Nielsen LH, Zor K, Boisen A, Christensen MV, Berthelsen J et al. Cellular effects and delivery propensity of penetratin is influenced by conjugation to parathyroid hormone fragment 1-34 in synergy with pH. Bioconjugate Chemistry. 2018;29(2):371-381. https://doi.org/10.1021/acs.bioconjchem.7b00687

Author

Kristensen, Mie ; Nielsen, Line Hagner ; Zor, Kinga ; Boisen, Anja ; Christensen, Malene Vinther ; Berthelsen, Jens ; Nielsen, Hanne Mørck. / Cellular effects and delivery propensity of penetratin is influenced by conjugation to parathyroid hormone fragment 1-34 in synergy with pH. In: Bioconjugate Chemistry. 2018 ; Vol. 29, No. 2. pp. 371-381.

Bibtex

@article{ae3c419b2361447091180552e1da6715,
title = "Cellular effects and delivery propensity of penetratin is influenced by conjugation to parathyroid hormone fragment 1-34 in synergy with pH",
abstract = "The cell-penetrating peptide (CPP) penetratin, has demonstrated potential as a carrier for transepithelial delivery of cargo peptides, such as the therapeutically relevant part of parathyroid hormone, i.e. PTH(1-34). The purpose of the present study was to elucidate the relevance of modifying the pH for PTH(1-34)-penetratin conjugates and for co-administered penetratin with PTH(1-34) in terms of transepithelial permeation of PTH(1-34) and cellular effects. Transepithelial permeation was assessed using monolayers of the Caco-2 cell culture model, and effects on Caco-2 cellular viability kinetics were evaluated by using the Real-Time-GLO assay as well as by microscopy following Tryphan blue staining. Morphological Caco-2 cell changes were studied exploiting the impedance-based xCELLigence system as well as optically using the oCelloscope setup. Finally, the effect of pH on the folding propensity of the PTH(1-34)-penetratin conjugate and its ability to disrupt lipid membranes were assessed by circular dichroism (CD) spectroscopy and the calcein release assay, respectively. The transepithelial PTH(1-34) permeation was not pH-dependent when applying the co-administration approach. However, by applying the conjugation approach, the PTH(1-34) permeation was significantly enhanced by lowering the pH from 7.4 to 5, but also associated with a compromised barrier and a lowering of the cellular viability. The negative effects on the cellular viability following cellular incubation with the PTH(1-34)-penetratin conjugate were moreover confirmed during real-time monitoring of the Caco-2 cell viability as well as by enhanced Tryphan blue uptake. In addition, morphological changes were primarily observed for cells incubated with the PTH(1-34)-penetratin conjugate at pH 5, which was moreover demonstrated to have an enhanced membrane permeating effect following lowering of the pH from 7.4 to 5. The latter observation was, however, not a result of better secondary folding propensity at pH 5 when compared to pH 7.4.",
author = "Mie Kristensen and Nielsen, {Line Hagner} and Kinga Zor and Anja Boisen and Christensen, {Malene Vinther} and Jens Berthelsen and Nielsen, {Hanne M{\o}rck}",
year = "2018",
doi = "10.1021/acs.bioconjchem.7b00687",
language = "English",
volume = "29",
pages = "371--381",
journal = "Bioconjugate Chemistry",
issn = "1043-1802",
publisher = "American Chemical Society",
number = "2",

}

RIS

TY - JOUR

T1 - Cellular effects and delivery propensity of penetratin is influenced by conjugation to parathyroid hormone fragment 1-34 in synergy with pH

AU - Kristensen, Mie

AU - Nielsen, Line Hagner

AU - Zor, Kinga

AU - Boisen, Anja

AU - Christensen, Malene Vinther

AU - Berthelsen, Jens

AU - Nielsen, Hanne Mørck

PY - 2018

Y1 - 2018

N2 - The cell-penetrating peptide (CPP) penetratin, has demonstrated potential as a carrier for transepithelial delivery of cargo peptides, such as the therapeutically relevant part of parathyroid hormone, i.e. PTH(1-34). The purpose of the present study was to elucidate the relevance of modifying the pH for PTH(1-34)-penetratin conjugates and for co-administered penetratin with PTH(1-34) in terms of transepithelial permeation of PTH(1-34) and cellular effects. Transepithelial permeation was assessed using monolayers of the Caco-2 cell culture model, and effects on Caco-2 cellular viability kinetics were evaluated by using the Real-Time-GLO assay as well as by microscopy following Tryphan blue staining. Morphological Caco-2 cell changes were studied exploiting the impedance-based xCELLigence system as well as optically using the oCelloscope setup. Finally, the effect of pH on the folding propensity of the PTH(1-34)-penetratin conjugate and its ability to disrupt lipid membranes were assessed by circular dichroism (CD) spectroscopy and the calcein release assay, respectively. The transepithelial PTH(1-34) permeation was not pH-dependent when applying the co-administration approach. However, by applying the conjugation approach, the PTH(1-34) permeation was significantly enhanced by lowering the pH from 7.4 to 5, but also associated with a compromised barrier and a lowering of the cellular viability. The negative effects on the cellular viability following cellular incubation with the PTH(1-34)-penetratin conjugate were moreover confirmed during real-time monitoring of the Caco-2 cell viability as well as by enhanced Tryphan blue uptake. In addition, morphological changes were primarily observed for cells incubated with the PTH(1-34)-penetratin conjugate at pH 5, which was moreover demonstrated to have an enhanced membrane permeating effect following lowering of the pH from 7.4 to 5. The latter observation was, however, not a result of better secondary folding propensity at pH 5 when compared to pH 7.4.

AB - The cell-penetrating peptide (CPP) penetratin, has demonstrated potential as a carrier for transepithelial delivery of cargo peptides, such as the therapeutically relevant part of parathyroid hormone, i.e. PTH(1-34). The purpose of the present study was to elucidate the relevance of modifying the pH for PTH(1-34)-penetratin conjugates and for co-administered penetratin with PTH(1-34) in terms of transepithelial permeation of PTH(1-34) and cellular effects. Transepithelial permeation was assessed using monolayers of the Caco-2 cell culture model, and effects on Caco-2 cellular viability kinetics were evaluated by using the Real-Time-GLO assay as well as by microscopy following Tryphan blue staining. Morphological Caco-2 cell changes were studied exploiting the impedance-based xCELLigence system as well as optically using the oCelloscope setup. Finally, the effect of pH on the folding propensity of the PTH(1-34)-penetratin conjugate and its ability to disrupt lipid membranes were assessed by circular dichroism (CD) spectroscopy and the calcein release assay, respectively. The transepithelial PTH(1-34) permeation was not pH-dependent when applying the co-administration approach. However, by applying the conjugation approach, the PTH(1-34) permeation was significantly enhanced by lowering the pH from 7.4 to 5, but also associated with a compromised barrier and a lowering of the cellular viability. The negative effects on the cellular viability following cellular incubation with the PTH(1-34)-penetratin conjugate were moreover confirmed during real-time monitoring of the Caco-2 cell viability as well as by enhanced Tryphan blue uptake. In addition, morphological changes were primarily observed for cells incubated with the PTH(1-34)-penetratin conjugate at pH 5, which was moreover demonstrated to have an enhanced membrane permeating effect following lowering of the pH from 7.4 to 5. The latter observation was, however, not a result of better secondary folding propensity at pH 5 when compared to pH 7.4.

U2 - 10.1021/acs.bioconjchem.7b00687

DO - 10.1021/acs.bioconjchem.7b00687

M3 - Journal article

C2 - 29155563

VL - 29

SP - 371

EP - 381

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

SN - 1043-1802

IS - 2

ER -

ID: 186908726