PCSK9 facilitates melanoma pathogenesis via a network regulating tumor immunity

Yan Gu; Xiaozeng Lin; Ying Dong; Geoffrey Wood; Nabil G Seidah; Geoff Werstuck; Pierre Major; Michael Bonert; Anil Kapoor; Damu Tang

doi:10.1186/s13046-022-02584-y

PCSK9 facilitates melanoma pathogenesis via a network regulating tumor immunity

J Exp Clin Cancer Res. 2023 Jan 2;42(1):2. doi: 10.1186/s13046-022-02584-y.

Authors

Yan Gu^{1

2

3}, Xiaozeng Lin^{1

2

3}, Ying Dong^{1

2

3}, Geoffrey Wood⁴, Nabil G Seidah⁵, Geoff Werstuck⁶, Pierre Major⁷, Michael Bonert^{3

8}, Anil Kapoor^{9

10

11}, Damu Tang^{12

13

14}

Affiliations

¹ Urological Cancer Center for Research and Innovation (UCCRI), T3310, St. Joseph's Hospital, 50 Charlton Ave East, Hamilton, ON, L8N 4A6, Canada.
² Department of Surgery, McMaster University, Hamilton, ON, L8S 4K1, Canada.
³ The Research Institute of St Joe's Hamilton, G344, St. Joseph's Hospital, Hamilton, ON, L8N 4A6, Canada.
⁴ Department of Pathology, University of Guelph, Guelph, ON, N1G 2W1, Canada.
⁵ Laboratory of Biochemical Neuroendocrinology, Montreal Clinical Research Institute, University of Montreal, Montreal, QC, H2W 1R7, Canada.
⁶ Department of Medicine, McMaster University, Hamilton, ON, L8S 4K1, Canada.
⁷ Department of Oncology, McMaster University, Hamilton, ON, L8S 4K1, Canada.
⁸ Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, L8S 4K1, Canada.
⁹ Urological Cancer Center for Research and Innovation (UCCRI), T3310, St. Joseph's Hospital, 50 Charlton Ave East, Hamilton, ON, L8N 4A6, Canada. akapoor@mcmaster.ca.
¹⁰ Department of Surgery, McMaster University, Hamilton, ON, L8S 4K1, Canada. akapoor@mcmaster.ca.
¹¹ The Research Institute of St Joe's Hamilton, G344, St. Joseph's Hospital, Hamilton, ON, L8N 4A6, Canada. akapoor@mcmaster.ca.
¹² Urological Cancer Center for Research and Innovation (UCCRI), T3310, St. Joseph's Hospital, 50 Charlton Ave East, Hamilton, ON, L8N 4A6, Canada. damut@mcmaster.ca.
¹³ Department of Surgery, McMaster University, Hamilton, ON, L8S 4K1, Canada. damut@mcmaster.ca.
¹⁴ The Research Institute of St Joe's Hamilton, G344, St. Joseph's Hospital, Hamilton, ON, L8N 4A6, Canada. damut@mcmaster.ca.

Abstract

Background: PCSK9 regulates cholesterol homeostasis and promotes tumorigenesis. However, the relevance of these two actions and the mechanisms underlying PCSK9's oncogenic roles in melanoma and other cancers remain unclear.

Methods: PCSK9's association with melanoma was analysed using the TCGA dataset. Empty vector (EV), PCSK9, gain-of-function (D374Y), and loss-of-function (Q152H) PCSK9 mutant were stably-expressed in murine melanoma B16 cells and studied for impact on B16 cell-derived oncogenesis in vitro and in vivo using syngeneic C57BL/6 and Pcsk9^-/- mice. Intratumoral accumulation of cholesterol was determined. RNA-seq was performed on individual tumor types. Differentially-expressed genes (DEGs) were derived from the comparisons of B16 PCSK9, B16 D374Y, or B16 Q152H tumors to B16 EV allografts and analysed for pathway alterations.

Results: PCSK9 expression and its network negatively correlated with the survival probability of patients with melanoma. PCSK9 promoted B16 cell proliferation, migration, and growth in soft agar in vitro, formation of tumors in C57BL/6 mice in vivo, and accumulation of intratumoral cholesterol in a manner reflecting its regulation of the low-density lipoprotein receptor (LDLR): Q152H, EV, PCSK9, and D374Y. Tumor-associated T cells, CD8 + T cells, and NK cells were significantly increased in D374Y tumors along with upregulations of multiple immune checkpoints, IFNγ, and 143 genes associated with T cell dysfunction. Overlap of 36 genes between the D374Y DEGs and the PCSK9 DEGs predicted poor prognosis of melanoma and resistance to immune checkpoint blockade (ICB) therapy. CYTH4, DENND1C, AOAH, TBC1D10C, EPSTI1, GIMAP7, and FASL (FAS ligand) were novel predictors of ICB therapy and displayed high level of correlations with multiple immune checkpoints in melanoma and across 30 human cancers. We observed FAS ligand being among the most robust biomarkers of ICB treatment and constructed two novel and effective multigene panels predicting response to ICB therapy. The profiles of allografts produced by B16 EV, PCSK9, D374Y, and Q152H remained comparable in C57BL/6 and Pcsk9^-/- mice.

Conclusions: Tumor-derived PCSK9 plays a critical role in melanoma pathogenesis. PCSK9's oncogenic actions are associated with intratumoral cholesterol accumulation. PCSK9 systemically affects the immune system, contributing to melanoma immune evasion. Novel biomarkers derived from the PCSK9-network effectively predicted ICB therapy responses.

Keywords: Biomarkers of ICB; Immune checkpoint blockade (ICB); Melanoma; PCSK9; Tumorigenesis.

MeSH terms

Animals
Cell Adhesion Molecules
Fas Ligand Protein
GTPase-Activating Proteins
Guanine Nucleotide Exchange Factors
Humans
Melanoma* / genetics
Melanoma, Experimental* / genetics
Melanoma, Experimental* / pathology
Mice
Mice, Inbred C57BL
Proprotein Convertase 9 / genetics

Substances

PCSK9 protein, human
Proprotein Convertase 9
Fas Ligand Protein
CYTH4 protein, human
Cell Adhesion Molecules
Guanine Nucleotide Exchange Factors
Pcsk9 protein, mouse
TBC1D10C protein, mouse
GTPase-Activating Proteins