Overcome Chemoresistance: Biophysical and Structural Analysis of Synthetic FHIT-Derived Peptides.

Maria Carmina Scala,Delia Lanzillotta,Francesco Trapasso,Simona Musella,Stella Maria Cascioferro,Marina Sala,Pietro Campiglia,Giuseppe Bifulco,Simone Di Micco,Veronica Di Sarno,Barbara Parrino

doi:10.3389/fmolb.2021.715263

Abstract

The fragile histidine triad (FHIT) protein is a member of the large and ubiquitous histidine triad (HIT) family of proteins. On the basis of genetic evidence, it has been postulated that the FHIT protein may function as tumor suppressor, implying a role for the FHIT protein in carcinogenesis. Recently, Gaudio et al. reported that FHIT binds and delocalizes annexin A4 (ANXA4) from plasma membrane to cytosol in paclitaxel-resistant lung cancer cells, thus restoring their chemosensitivity to the drug. They also identified the smallest protein sequence of the FHIT still interacting with ANXA4, ranging from position 7 to 13: QHLIKPS. This short sequence of FHIT protein was not only able to bind ANXA4 but also to hold its target in the cytosol during paclitaxel treatment, thus avoiding ANXA4 translocation to the inner side of the cell membrane. Starting from these results, to obtain much information about structure requirements involved in the interaction of the peptide mentioned above, we synthetized a panel of seven peptides through an Ala-scan approach. In detail, to study the binding of FHIT derived peptides with ANXA4, we applied a combination of different biophysical techniques such as differential scanning fluorimetry (DSF), surface plasmon resonance (SPR), and microscale thermophoresis (MST). Circular dichroism (CD) and nuclear magnetic resonance (NMR) were used to determine the conformational structure of the lead peptide (7–13) and peptides generated from ala-scan technique. The application of different biophysical and structural techniques, integrated by a preliminary biological evaluation, allowed us to build a solid structure activity relationship on the synthesized peptides.

Highlights

Chemoresistance is the ability of cancer cells to evade the action of several classes of anti-cancer drugs, becoming a serious problem that cancer research seeks to understand and overcome.The molecular mechanisms of how cancer cells promote their own survival and avoid apoptosis in response to commonly used chemotherapeutics are multiple and consist of a set of signaling pathways, which can be activated by a large amount of stimuli to promote chemoresistance (Housman et al, 2014; Bukowski et al, 2020)
The molecular mapping of the Fhit protein domain that interacts with Annexin A4 led to the restriction of the interaction domain to amino acids 7 to 13 of the Fhit protein
Fhit peptide retains the property of the native protein in inhibiting Annexin A4 translocation from cytosol to plasma membrane in A549 and Calu-2 lung cancer cells treated with paclitaxel (Gaudio et al, 2013)

Summary

Introduction

Chemoresistance is the ability of cancer cells to evade the action of several classes of anti-cancer drugs, becoming a serious problem that cancer research seeks to understand and overcome. The molecular mechanisms of how cancer cells promote their own survival and avoid apoptosis in response to commonly used chemotherapeutics are multiple and consist of a set of signaling pathways, which can be activated by a large amount of stimuli to promote chemoresistance (Housman et al, 2014; Bukowski et al, 2020). In this regard, ANXA4 is an interesting.

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Results

Conclusion