Exogenous Cripto-1 Suppresses Self-Renewal of Cancer Stem Cell Model

Md Jahangir Alam, Ryota Takahashi, Said M. Afify, Aung Ko Ko Oo, Kazuki Kumon, Hend M. Nawara, Aprilliana Cahya Khayrani, Juan Du, Hussein Maram, Akimasa Seno, David S. Salomon, Masaharu Seno

Research output: Contribution to journalArticle

Abstract

Cripto-1 is a glycophosphatidylinositol (GPI) anchored signaling protein of epidermal growth factor (EGF)-Cripto-1-FRL1-Cryptic (CFC) family and plays a significant role in the early developmental stages and in the different types of cancer cells, epithelial to mesenchymal transition and tumor angiogenesis. Previously, we have developed cancer stem cells (miPS-LLCcm) from mouse iPSCs by culturing them in the presence of conditioned medium of Lewis Lung Carcinoma (LLC) cells for four weeks. Nodal and Cripto-1 were confirmed to be expressed in miPS-LLCcm cells by quantitative reverse transcription PCR (rt-qPCR) implying that Cr-1 was required in maintaining stemness. To investigate the biological effect of adding exogenous soluble CR-1 to the cancer stem cells, we have prepared a C-terminally truncated soluble form of recombinant human CR-1 protein (rhsfCR-1), in which the GPI anchored moiety was removed by substitution of a stop codon through site-directed mutagenesis. rhsfCR-1 effectively suppressed the proliferation and sphere forming ability of miPS-LLCcm cells in a dose-dependent manner in the range of 0 to 5 µg/mL, due to the suppression of Nodal-Cripto-1/ALK4/Smad2 signaling pathway. Frequency of sphere-forming cells was dropped from 1/40 to 1/69 by rhsfCR-1 at 1 µg/mL. Moreover, rhsfCR-1 in the range of 0 to 1 µg/mL also limited the differentiation of miPS-LLCcm cells into vascular endothelial cells probably due to the suppression of self-renewal, which should reduce the number of cells with stemness property. As demonstrated by a soluble form of exogenous Cripto-1 in this study, the efficient blockade would be an attractive way to study Cripto-1 dependent cancer stem cell properties for therapeutic application.

Original languageEnglish
JournalInternational Journal of Molecular Sciences
Volume19
Issue number11
DOIs
Publication statusPublished - Oct 26 2018

Fingerprint

Neoplastic Stem Cells
stem cells
Stem cells
cancer
cells
Cells
Proteins
Mutagenesis
Endothelial cells
Transcription
Conditioned Culture Medium
Epidermal Growth Factor
retarding
Tumors
proteins
Substitution reactions
mutagenesis
angiogenesis
biological effects
Lewis Lung Carcinoma

Keywords

  • cancer stem cells
  • Cripto-1
  • miPS-LLCcm
  • mouse iPS
  • recombinant Cripto-1
  • self-renewal

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

Cite this

Exogenous Cripto-1 Suppresses Self-Renewal of Cancer Stem Cell Model. / Alam, Md Jahangir; Takahashi, Ryota; Afify, Said M.; Oo, Aung Ko Ko; Kumon, Kazuki; Nawara, Hend M.; Khayrani, Aprilliana Cahya; Du, Juan; Maram, Hussein; Seno, Akimasa; Salomon, David S.; Seno, Masaharu.

In: International Journal of Molecular Sciences, Vol. 19, No. 11, 26.10.2018.

Research output: Contribution to journalArticle

Alam, MJ, Takahashi, R, Afify, SM, Oo, AKK, Kumon, K, Nawara, HM, Khayrani, AC, Du, J, Maram, H, Seno, A, Salomon, DS & Seno, M 2018, 'Exogenous Cripto-1 Suppresses Self-Renewal of Cancer Stem Cell Model', International Journal of Molecular Sciences, vol. 19, no. 11. https://doi.org/10.3390/ijms19113345
Alam, Md Jahangir ; Takahashi, Ryota ; Afify, Said M. ; Oo, Aung Ko Ko ; Kumon, Kazuki ; Nawara, Hend M. ; Khayrani, Aprilliana Cahya ; Du, Juan ; Maram, Hussein ; Seno, Akimasa ; Salomon, David S. ; Seno, Masaharu. / Exogenous Cripto-1 Suppresses Self-Renewal of Cancer Stem Cell Model. In: International Journal of Molecular Sciences. 2018 ; Vol. 19, No. 11.
@article{fd01cb1e8e934516b34580d7f036151c,
title = "Exogenous Cripto-1 Suppresses Self-Renewal of Cancer Stem Cell Model",
abstract = "Cripto-1 is a glycophosphatidylinositol (GPI) anchored signaling protein of epidermal growth factor (EGF)-Cripto-1-FRL1-Cryptic (CFC) family and plays a significant role in the early developmental stages and in the different types of cancer cells, epithelial to mesenchymal transition and tumor angiogenesis. Previously, we have developed cancer stem cells (miPS-LLCcm) from mouse iPSCs by culturing them in the presence of conditioned medium of Lewis Lung Carcinoma (LLC) cells for four weeks. Nodal and Cripto-1 were confirmed to be expressed in miPS-LLCcm cells by quantitative reverse transcription PCR (rt-qPCR) implying that Cr-1 was required in maintaining stemness. To investigate the biological effect of adding exogenous soluble CR-1 to the cancer stem cells, we have prepared a C-terminally truncated soluble form of recombinant human CR-1 protein (rhsfCR-1), in which the GPI anchored moiety was removed by substitution of a stop codon through site-directed mutagenesis. rhsfCR-1 effectively suppressed the proliferation and sphere forming ability of miPS-LLCcm cells in a dose-dependent manner in the range of 0 to 5 µg/mL, due to the suppression of Nodal-Cripto-1/ALK4/Smad2 signaling pathway. Frequency of sphere-forming cells was dropped from 1/40 to 1/69 by rhsfCR-1 at 1 µg/mL. Moreover, rhsfCR-1 in the range of 0 to 1 µg/mL also limited the differentiation of miPS-LLCcm cells into vascular endothelial cells probably due to the suppression of self-renewal, which should reduce the number of cells with stemness property. As demonstrated by a soluble form of exogenous Cripto-1 in this study, the efficient blockade would be an attractive way to study Cripto-1 dependent cancer stem cell properties for therapeutic application.",
keywords = "cancer stem cells, Cripto-1, miPS-LLCcm, mouse iPS, recombinant Cripto-1, self-renewal",
author = "Alam, {Md Jahangir} and Ryota Takahashi and Afify, {Said M.} and Oo, {Aung Ko Ko} and Kazuki Kumon and Nawara, {Hend M.} and Khayrani, {Aprilliana Cahya} and Juan Du and Hussein Maram and Akimasa Seno and Salomon, {David S.} and Masaharu Seno",
year = "2018",
month = "10",
day = "26",
doi = "10.3390/ijms19113345",
language = "English",
volume = "19",
journal = "International Journal of Molecular Sciences",
issn = "1661-6596",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
number = "11",

}

TY - JOUR

T1 - Exogenous Cripto-1 Suppresses Self-Renewal of Cancer Stem Cell Model

AU - Alam, Md Jahangir

AU - Takahashi, Ryota

AU - Afify, Said M.

AU - Oo, Aung Ko Ko

AU - Kumon, Kazuki

AU - Nawara, Hend M.

AU - Khayrani, Aprilliana Cahya

AU - Du, Juan

AU - Maram, Hussein

AU - Seno, Akimasa

AU - Salomon, David S.

AU - Seno, Masaharu

PY - 2018/10/26

Y1 - 2018/10/26

N2 - Cripto-1 is a glycophosphatidylinositol (GPI) anchored signaling protein of epidermal growth factor (EGF)-Cripto-1-FRL1-Cryptic (CFC) family and plays a significant role in the early developmental stages and in the different types of cancer cells, epithelial to mesenchymal transition and tumor angiogenesis. Previously, we have developed cancer stem cells (miPS-LLCcm) from mouse iPSCs by culturing them in the presence of conditioned medium of Lewis Lung Carcinoma (LLC) cells for four weeks. Nodal and Cripto-1 were confirmed to be expressed in miPS-LLCcm cells by quantitative reverse transcription PCR (rt-qPCR) implying that Cr-1 was required in maintaining stemness. To investigate the biological effect of adding exogenous soluble CR-1 to the cancer stem cells, we have prepared a C-terminally truncated soluble form of recombinant human CR-1 protein (rhsfCR-1), in which the GPI anchored moiety was removed by substitution of a stop codon through site-directed mutagenesis. rhsfCR-1 effectively suppressed the proliferation and sphere forming ability of miPS-LLCcm cells in a dose-dependent manner in the range of 0 to 5 µg/mL, due to the suppression of Nodal-Cripto-1/ALK4/Smad2 signaling pathway. Frequency of sphere-forming cells was dropped from 1/40 to 1/69 by rhsfCR-1 at 1 µg/mL. Moreover, rhsfCR-1 in the range of 0 to 1 µg/mL also limited the differentiation of miPS-LLCcm cells into vascular endothelial cells probably due to the suppression of self-renewal, which should reduce the number of cells with stemness property. As demonstrated by a soluble form of exogenous Cripto-1 in this study, the efficient blockade would be an attractive way to study Cripto-1 dependent cancer stem cell properties for therapeutic application.

AB - Cripto-1 is a glycophosphatidylinositol (GPI) anchored signaling protein of epidermal growth factor (EGF)-Cripto-1-FRL1-Cryptic (CFC) family and plays a significant role in the early developmental stages and in the different types of cancer cells, epithelial to mesenchymal transition and tumor angiogenesis. Previously, we have developed cancer stem cells (miPS-LLCcm) from mouse iPSCs by culturing them in the presence of conditioned medium of Lewis Lung Carcinoma (LLC) cells for four weeks. Nodal and Cripto-1 were confirmed to be expressed in miPS-LLCcm cells by quantitative reverse transcription PCR (rt-qPCR) implying that Cr-1 was required in maintaining stemness. To investigate the biological effect of adding exogenous soluble CR-1 to the cancer stem cells, we have prepared a C-terminally truncated soluble form of recombinant human CR-1 protein (rhsfCR-1), in which the GPI anchored moiety was removed by substitution of a stop codon through site-directed mutagenesis. rhsfCR-1 effectively suppressed the proliferation and sphere forming ability of miPS-LLCcm cells in a dose-dependent manner in the range of 0 to 5 µg/mL, due to the suppression of Nodal-Cripto-1/ALK4/Smad2 signaling pathway. Frequency of sphere-forming cells was dropped from 1/40 to 1/69 by rhsfCR-1 at 1 µg/mL. Moreover, rhsfCR-1 in the range of 0 to 1 µg/mL also limited the differentiation of miPS-LLCcm cells into vascular endothelial cells probably due to the suppression of self-renewal, which should reduce the number of cells with stemness property. As demonstrated by a soluble form of exogenous Cripto-1 in this study, the efficient blockade would be an attractive way to study Cripto-1 dependent cancer stem cell properties for therapeutic application.

KW - cancer stem cells

KW - Cripto-1

KW - miPS-LLCcm

KW - mouse iPS

KW - recombinant Cripto-1

KW - self-renewal

UR - http://www.scopus.com/inward/record.url?scp=85055619636&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85055619636&partnerID=8YFLogxK

U2 - 10.3390/ijms19113345

DO - 10.3390/ijms19113345

M3 - Article

C2 - 30373174

AN - SCOPUS:85055619636

VL - 19

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

SN - 1661-6596

IS - 11

ER -