Lipid-sensors, enigmatic-orphan and orphan nuclear receptors as therapeutic targets in breast-cancer

Abstract

Breast-cancer is heterogeneous and consists of various groups with different biological characteristics. Innovative pharmacological approaches accounting for this heterogeneity are needed. The forty eight human Nuclear-Hormone-Receptors are ligand-dependent transcription-factors and are classified into Endocrine-Receptors, Adopted-Orphan-Receptors (Lipid-sensors and Enigmatic-Orphans) and Orphan-receptors. Nuclear-Receptors represent ideal targets for the design/synthesis of pharmacological ligands. We provide an overview of the literature available on the expression and potential role played by Lipid-sensors, Enigmatic-Orphans and Orphan-Receptors in breast-cancer. The data are complemented by an analysis of the expression levels of each selected Nuclear-Receptor in the PAM50 breast-cancer groups, following re-elaboration of the data publicly available. The major aim is to support the idea that some of the Nuclear-Receptors represent largely unexploited therapeutic-targets in breast-cancer treatment/chemo-prevention. On the basis of our analysis, we conclude that the Lipid-Sensors, NR1C3, NR1H2 and NR1H3 are likely to be onco-suppressors in breast-cancer. The Enigmatic-Orphans, NR1F1 NR2A1 and NR3B3 as well as the Orphan-Receptors, NR0B1, NR0B2, NR1D1, NR2F1, NR2F2 and NR4A3 exert a similar action. These Nuclear-Receptors represent candidates for the development of therapeutic strategies aimed at increasing their expression or activating them in tumor cells. The group of Nuclear-Receptors endowed with potential oncogenic properties consists of the Lipid-Sensors, NR1C2 and NR1I2, the Enigmatic-Orphans, NR1F3, NR3B1 and NR5A2, as well as the Orphan-Receptors, NR2E1, NR2E3 and NR6A1. These oncogenic Nuclear-Receptors should be targeted with selective antagonists, reverse-agonists or agents/strategies capable of reducing their expression in breast-cancer cells.

Keywords: breast cancer; chemo-prevention; drug targets; nuclear receptors; treatment.

Figure 1. Correspondence between the PAM50 and the traditional classification of breast-cancer

A. Correspondence between the immuno-histochemical and the molecular/transcriptomic classification of breast cancer. Mammary-tumors are divided in four groups according to the traditional classification based on the immuno-histochemical determination of the ERα, PR and HER2 molecular markers. For each of the four groups, the percentage of cases showing the six indicated transcriptomic phenotypes determined on the basis of a modification of the PAM50 fingerprint is illustrated. B. Expression of ERα (Upper circles) and HER2 (Lower circles) among the molecular/transcriptomic subtypes of breast cancer. Mammary tumors are split in six groups according the transcriptomic phenotype as detailed in A.. For each of the six groups the percentage of cases showing positivity to ERα is indicated. The results indicates that practically the totality of Luminal-A and Luminal-B tumors and the vast majority of the Normal-like ones are ER⁺. In contrast, the vast majority of Basal-like and Claudin-Low tumors, which are aggregated into the PAM50 Basal group, are ER⁻. The data were obtained from the following article: Rivenbark AG, O'Connor SM, Coleman WB. Molecular and cellular heterogeneity in breast cancer: challenges for personalized medicine. Am J Pathol (2013) 183: 1113-24.

Figure 2. Phylogenetic tree and schematic structure of the human nuclear steroid receptors

A. The panel illustrates an unrooted phylogenetic tree of the human nuclear steroid receptors. The phylogenetic tree was obtained following alignment of the amino acid sequences of the indicated nuclear receptors using the Clustal-omega algorithm (Sievers F, Wilm A, Dineen DG, Gibson TJ, Karplus K, Li W, Lopez R, McWilliam H, Remmert M, Söding J, Thompson JD, Higgins D. Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega Molecular Systems Biology 7 Article number: 539 doi:10.1038/msb.2011.75) B. The schematic structure of a typical nuclear receptor (NR) is shown in the upper diagram. The N-terminal A- and B-regions are responsible for the ligand-independent activation function (AF-1). The C-region contains the DNA-binding domain, while the unstructured D-region connects the C- and E-regions. The E-region containing the ligand-binding pocket is responsible for the ligand-dependent activation function (AF-2). The function of the C-terminal F-region is undefined. The tridimensional structure of the C- and E-regions contained in a representative NR (ERα) is illustrated in the lower part of the figure.

Figure 3. Expression of Lipid-sensors in normal mammary glands and breast-cancer tissue

The box-plots illustrate the expression of the indicated mRNAs belonging to the Lipid-sensors family of nuclear receptors (NRs) in normal mammary-glands (NG), all breast-cancers (BC) and Luminal-A (LA), Luminal-B (LB), HER2 (HR), Basal (BL) and Normal-like (NL) PAM50 mammary-tumors. Underneath each box-plot, the tables show significant differences in the mRNA expression levels of each NR between the indicated groups. The results were obtained from the data available in the TCGA (The Cancer Genome Atlas; http://cancergenome.nih.gov). Normalization, quantification and statistical analysis on raw sequencing counts was performed using the Limma/Voom (http://bioconductor.org) package in R statistical environment. * (adjusted p < 0.01), ** (adjusted p < 0.001), *** (adjusted p < 0.0001).

Figure 4. Expression of Enigmatic-Orphan receptors in normal mammary glands and breast-cancer tissue

The box-plots illustrate the expression of the indicated mRNAs belonging to the Enigmatic-Orphans family of nuclear receptors (NRs) in normal mammary-glands (NG), all breast-cancers (BC) and the Luminal-A (LA), Luminal-B (LB), HER2 (HR), Basal (BL) and Normal-like (NL) PAM50 mammary-tumors. Underneath each box-plot, the tables show significant differences in the mRNA expression levels of each NR between the indicated groups. The results were obtained from the data available in the TCGA (The Cancer Genome Atlas; http://cancergenome.nih.gov). Normalization, quantification and statistical analysis on raw sequencing counts was performed using the Limma/Voom (http://bioconductor.org) package in R statistical environment. * (adjusted p < 0.01), ** (adjusted p < 0.001), *** (adjusted p < 0.0001).

Figure 5. Expression of Orphan-Receptors in normal mammary glands and breast-cancer tissue

The box-plots illustrate the expression of the indicated mRNAs belonging to the Orphan-Receptors family of nuclear receptors (NRs) in normal mammary-glands (NG), all breast-cancers (BC) and the Luminal-A (LA), Luminal-B (LB), HER2 (HR), Basal (BL) and Normal-like (NL) PAM50 mammary-tumors. Underneath each box-plot, the tables show significant differences in the mRNA expression levels of each NR between the indicated groups. The results were obtained from the data available in the TCGA (The-Cancer-Genome-Atlas: http://cancergenome.nih.gov). Normalization, quantification and statistical analysis on raw sequencing counts was performed using the Limma/Voom (http://bioconductor.org) package in R statistical environment. * (adjusted p < 0.01), ** (adjusted p < 0.001), *** (adjusted p < 0.0001).