CD66+ Cells in Cervical Precancers Are Partially Differentiated Progenitors with Neoplastic Traits

Abstract

Cervical cancers, a malignancy associated with oncogenic papillomaviruses, remain a major disease burden in the absence of effective implementation of preventive strategies. CD66+ cells have previously been identified as a tumor propagating subset in cervical cancers. We investigated the existence, differentiation state and neoplastic potential of CD66+ cells in a pre-cancer cell line harboring HPV31b episomes. The gene expression profile of CD66High cells overlaps with differentiated keratinocytes, neoplastic mesenchymal transition, cells of the squamo-columnar junction and cervical cancer cell line derived spheroids. There is elevated expression of DNMT1, Notch1 and the viral gene product E1^E4, in CD66High cells. Thus CD66High cells, in the absence of differentiating signals, express higher levels of key regulators of keratinocytes stemness, differentiation and the viral life cycle respectively. We also find a striking association of neoplastic traits including migration, invasion and colony formation in soft agar with CD66High cells. These properties and a distinct G2M enriched cell cycle profile are conserved in cells from cervical cancers. Principally, using a precancerous cell line, we propose that CD66High cells have an intermediate differentiation state with a cellular milieu connected with both viral replication and neoplastic potential and validate some key features in pre-cancer lesions. Such pathophysiologically relevant systems for defining cellular changes in the early phases of the disease process provide both mechanistic insight and potential therapeutic strategies. Collectively, our data provides a rationale for exploring novel therapeutic targets in CD66+ sub-sets during cancer progression.

Human papillomavirus 16E6 and NFX1-123 potentiate notch signaling and differentiation without activating cellular arrest

“…Recent studies support our findings and have found increased Notch1 expression co-occurring with both an increased growth potential and differentiation marker expression in cervical cancer cell lines as well as in HFKs expressing HPV 31 (Pattabiraman, Hong et al, 2014). Additionally, the parsing of differentiation and growth arrest pathways has been seen in the context of 16E7 expression (Lefort, Mandinova et al, 2007; Muller, Alunni-Fabbroni et al, 1999; Nguyen, Lefort et al, 2006), so there is precedence for an HPV oncoprotein uncoupling differentiation from cellular growth.…”

CD66 and CD49f expressing cells are associated with distinct neoplastic phenotypes and progression in human cervical cancer

“…2 show that both in primary tissue and cervical cancer cell line model, the leading CD49f positive cells are more basal and proliferative; whereas trailing CD66 cells are more differentiated, migratory and show features of EMT. We have previously shown [7] using dye-dilution experiments that in whole CaSki-spheroids, CD66À CD49fþcells proliferate more compared to other subsets. Also we have shown [7] that CD66 and CD49f subsets follow a hierarchy in the expression of differentiation markers S100P and Involucrin in sorted cells from cell line and patient samples.…”

“…We have previously shown [7] using dye-dilution experiments that in whole CaSki-spheroids, CD66À CD49fþcells proliferate more compared to other subsets. Also we have shown [7] that CD66 and CD49f subsets follow a hierarchy in the expression of differentiation markers S100P and Involucrin in sorted cells from cell line and patient samples. CD66þCD49fÀ cells express highest levels of S100P and involucrin, followed by CD66þCD49fþ cells and then the CD66ÀCD49fþve cells.…”

“…The CD66 gene is a part of the carcinoembryonic antigen related cell adhesion molecule (CEACAM) family with various splice variants, amongst which CD66a, c and e are highly expressed in several cancers and have been suggested to mediate poor outcomes in some instances [2e5]. We have recently characterised a subset of CD66þve cells in cancers [6] and precancers [7] of cervix with distinctive tumourigenic properties. While invasiveness of CD66þ cells in both cervical pre-cancer and cancers seems a hallmark of this subset, there are parallel complex observations on the differentiation and cell cycle status of this subset which has made the generation of simplistic models of tumourigenic progression somewhat difficult.…”

Immune ligands for cytotoxic T Lymphocytes CTLS in cancer stem cells CSCS

“…A monoclonal antibody against CEACAM1 enhanced the effects of melanoma reactive lymphocytes in a mouse model of human melanoma xenograft (91). Precancerous lesions of the uterine cervix that harbor HPV genetic elements co-express CEACAM1, allowing for viral replication (92). Thus all TIM-3 ligands have multiple functions, besides inhibiting CTLs that affect various aspects of carcinogenesis.…”