Peritoneal carcinomatosis and peritoneal sarcomatosis (PC/PS) is a
potential complication of nearly all solid tumors and results in profoundly
increased morbidity and mortality. Despite the ubiquity of PC/PS, there are no
clinically relevant targeted therapies for either its treatment or prevention.
To identify potential therapies, we developed in vitro models
of PC/PS using tumor cell lines and patient-derived spheroids (PDS) that
recapitulate anoikis resistance and spheroid proliferation across multiple
cancer types. Epithelial- and mesenchymal-derived cancer cell lines (YOU, PANC1,
HEYA8, CHLA10, TC71) were used to generate spheroids and establish growth
characteristics. Differential gene expression analyses of these spheroids to
matched adherent cells, revealed a consensus spheroid signature. This spheroid
signature discriminates primary tumor specimens from tumor cells found in
ascites of ovarian cancer patients and in our PDS models. Key in this gene
expression signature is BNIP3 and BNIP3L, known regulators of autophagy and
apoptosis. Elevated BNIP3 mRNA expression is associated with poor survival in
ovarian cancer patients and elevated BNIP3 protein, as measured by
immunohistochemistry (IHC), is also associated with higher-grade tumors and
shorter survival. Pharmacological induction of autophagy with rapamycin
significantly increased spheroid formation and survival while decreasing the
induction of apoptosis. In contrast, the autophagy inhibitor hydroxychloroquine
abrogated spheroid formation with a clear increase in apoptosis. Modulation of
BNIP3 and the critical autophagy gene Beclin-1 (BECN1) also caused a significant
decrease in spheroid formation. Combined, these data demonstrate how modulation
of BNIP3-related autophagy, in patient-derived spheroids and in
vitro spheroid models, alters the survival and morphology of
spheroids.