Multi-stage inhibition in breast cancer metastasis by orally active triple conjugate, LHTD4 (low molecular weight heparin-taurocholate-tetrameric deoxycholate)

Farzana Alam, Taslim A. Al-Hilal, Jooho Park, Jeong Uk Choi, Foyez Mahmud, Jee Heon Jeong, In-San Kim, Sang Yoon Kim, Seung Rim Hwang, Youngro Byun

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Targeting multiple stages in metastatic breast cancer is one of the effective ways to inhibit metastatic progression. To target human metastatic breast cancer as well as improving patient compliance, we developed an orally active low molecular weight heparin (LMWH)-taurocholate conjugated with tetrameric deoxycholic acid, namely LHTD4, which followed by physical complexation with a synthetic bile acid enhancer, DCK. In breast cancer, both transforming growth factor-β1 (TGF-β1) and CXCL12 exhibit enhanced metastatic activity during the initiation and progression stages of breast cancer, thus we direct the focus on investigating the antimetastatic effect of LHTD4/DCK complex by targeting TGF-β1 and CXCL12. Computer simulation study and SPR analysis were performed for the binding confirmation of LHTD4 with TGF-β1 and CXCL12. We carried out in vitro phosphorylation assays of the consecutive receptors of TGF-β1 and CXCL12 (TGF-β1R1 and CXCR4, respectively). Effects of LHTD4 on in vitro cell migration (induced by TGF-β1) and chemotaxis (mediated by CXCL12) were investigated. The in vivo anti-metastatic effect of LHTD4 was evaluated in an accelerated metastasis model and an orthotopic MDA-MB-231 breast cancer model. The obtained KD values of TGF-β1 and CXCL12 with LHTD4 were 0.85 and 0.019 μM respectively. The simulation study showed that binding affinities of LHTD4 fragment with either TGF-β1 or CXCL12 through additional electrostatic interaction was more stable than that of LMWH fragment. In vitro phosphorylation assays of TGF-β1R1 and CXCR4 showed that the effective inhibition of receptor phosphorylation was observed with the treatment of LHTD4. The expressions of epithelial to mesenchymal transition (EMT) marker proteins such as vimentin and Snail were prevented by LTHD4 treatment in in vitro studies with TGF-β1 treated MDA-MB-231 cells. Moreover, we observed that LHTD4 negatively regulated the functions of TGF-β1 and CXCL12 on migration and invasion of breast cancer cell. In several advanced orthotopic and experimental breast cancer metastasis murine models, the treatment with LHTD4 (5 mg/kg daily, p.o.) significantly inhibited metastasis compared to the control. Overall, LHTD4 exhibited anti-metastatic effects by inhibiting TGF-β1 and CXCL12, and the clinically relevant dose of orally active LHTD4 was found to be effective in preclinical studies without any apparent toxicity.

Original languageEnglish
Pages (from-to)56-67
Number of pages12
JournalBiomaterials
Volume86
DOIs
Publication statusPublished - 2016 Apr 1
Externally publishedYes

Fingerprint

Taurocholic Acid
Deoxycholic Acid
Low Molecular Weight Heparin
Transforming Growth Factors
Molecular weight
Breast Neoplasms
Neoplasm Metastasis
Phosphorylation
Assays
Inhibition (Psychology)
Intercellular Signaling Peptides and Proteins
Heparin
Epithelial-Mesenchymal Transition
Acids
Vimentin
Chemotaxis
Patient Compliance
Coulomb interactions
Complexation
Bile Acids and Salts

Keywords

  • CXCL12
  • Low molecular weight heparin
  • Metastasis
  • Multi-stage targeting
  • Polymeric bile acid
  • TGF-β1

ASJC Scopus subject areas

  • Bioengineering
  • Ceramics and Composites
  • Biophysics
  • Biomaterials
  • Mechanics of Materials

Cite this

Multi-stage inhibition in breast cancer metastasis by orally active triple conjugate, LHTD4 (low molecular weight heparin-taurocholate-tetrameric deoxycholate). / Alam, Farzana; Al-Hilal, Taslim A.; Park, Jooho; Choi, Jeong Uk; Mahmud, Foyez; Jeong, Jee Heon; Kim, In-San; Kim, Sang Yoon; Hwang, Seung Rim; Byun, Youngro.

In: Biomaterials, Vol. 86, 01.04.2016, p. 56-67.

Research output: Contribution to journalArticle

Alam, Farzana ; Al-Hilal, Taslim A. ; Park, Jooho ; Choi, Jeong Uk ; Mahmud, Foyez ; Jeong, Jee Heon ; Kim, In-San ; Kim, Sang Yoon ; Hwang, Seung Rim ; Byun, Youngro. / Multi-stage inhibition in breast cancer metastasis by orally active triple conjugate, LHTD4 (low molecular weight heparin-taurocholate-tetrameric deoxycholate). In: Biomaterials. 2016 ; Vol. 86. pp. 56-67.
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AU - Al-Hilal, Taslim A.

AU - Park, Jooho

AU - Choi, Jeong Uk

AU - Mahmud, Foyez

AU - Jeong, Jee Heon

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AU - Byun, Youngro

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