TY - JOUR
T1 - Pancreatic Tumor Microenvironment Factor Promotes Cancer Stemness via SPP1–CD44 Axis
AU - Nallasamy, Palanisamy
AU - Nimmakayala, Rama Krishna
AU - Karmakar, Saswati
AU - Leon, Frank
AU - Seshacharyulu, Parthasarathy
AU - Lakshmanan, Imayavaramban
AU - Rachagani, Satyanarayana
AU - Mallya, Kavita
AU - Zhang, Chunmeng
AU - Ly, Quan P.
AU - Myers, Molly S.
AU - Josh, Lindenberger
AU - Grabow, Corinn E.
AU - Gautam, Shailendra K.
AU - Kumar, Sushil
AU - Lele, Subodh M.
AU - Jain, Maneesh
AU - Batra, Surinder K.
AU - Ponnusamy, Moorthy P.
N1 - Funding Information:
Funding The authors and work in this article were supported, in part, by the following grants from the National Institutes of Health (P01 CA217798, R01 CA210637, R01 CA183459, R01 CA195586, R01 CA201444, R01 CA228524, U01 CA20046, and U01 CA210240) and the Nebraska Department of Health and Human Services (LB595).
Publisher Copyright:
© 2021
PY - 2021/12
Y1 - 2021/12
N2 - Background & Aims: Tumor-microenvironment factors and cancer stem cells (CSCs) play a critical role in the aggressiveness of pancreatic cancer (PC). However, the degree to which tumor-microenvironment factors promote stemness remains unexplored. Here, we examined whether cancer-associated fibroblasts (CAFs) promote CSC features in PC. Methods: PC cells were treated long-term (30, 60, and 90 days) with conditioned media (CM)-derived from normal human fibroblasts (NFs) and CAFs. The stemness features of tumorsphere formation and stemness populations, along with CSCs markers, were analyzed using 2-dimensional and 3-dimensional sodium alginate bead-based co-culture models. Immunohistochemistry and immunofluorescence staining were performed for CSCs and fibroblast markers in autochthonous KrasG12D/+; Trp53R172H/+; Pdx1-Cre mice and human pancreatic tumors. Polymerase chain reaction array and gene knockdown were performed to identify the mechanism of stemness enrichment. Results: Long-term treatment of PC cells with CAF-CM enriched stemness, as indicated by significantly higher CD44+, ALDH+, and AF+ populations in PC cells. Increased tumorsphere formation and elevated CSC, self-renewal, and drug-resistance markers in CAF-CM–treated PC cells were observed. In addition, CAFs co-cultured with PC cells in the 3-dimensional model showed a substantial increase in stemness features. CD44 and α–smooth muscle actin were positively correlated and their expressions progressively increased from the early to late stages of KrasG12D/+; Trp53R172H/+; Pdx1-Cre mouse and human pancreatic tumors. Osteopontin/secreted phosphoprotein 1 was identified as the top differentially overexpressed gene in CAF-CM–treated PC cells and knockdown of osteopontin/secreted phosphoprotein 1 significantly reduced stemness characteristics in CAF-CM–treated PC cells. Conclusions: Our data uncovered novel insight into the interplay between CAF and enrichment of stemness population through the osteopontin/secreted phosphoprotein 1–CD44 axis in PC.
AB - Background & Aims: Tumor-microenvironment factors and cancer stem cells (CSCs) play a critical role in the aggressiveness of pancreatic cancer (PC). However, the degree to which tumor-microenvironment factors promote stemness remains unexplored. Here, we examined whether cancer-associated fibroblasts (CAFs) promote CSC features in PC. Methods: PC cells were treated long-term (30, 60, and 90 days) with conditioned media (CM)-derived from normal human fibroblasts (NFs) and CAFs. The stemness features of tumorsphere formation and stemness populations, along with CSCs markers, were analyzed using 2-dimensional and 3-dimensional sodium alginate bead-based co-culture models. Immunohistochemistry and immunofluorescence staining were performed for CSCs and fibroblast markers in autochthonous KrasG12D/+; Trp53R172H/+; Pdx1-Cre mice and human pancreatic tumors. Polymerase chain reaction array and gene knockdown were performed to identify the mechanism of stemness enrichment. Results: Long-term treatment of PC cells with CAF-CM enriched stemness, as indicated by significantly higher CD44+, ALDH+, and AF+ populations in PC cells. Increased tumorsphere formation and elevated CSC, self-renewal, and drug-resistance markers in CAF-CM–treated PC cells were observed. In addition, CAFs co-cultured with PC cells in the 3-dimensional model showed a substantial increase in stemness features. CD44 and α–smooth muscle actin were positively correlated and their expressions progressively increased from the early to late stages of KrasG12D/+; Trp53R172H/+; Pdx1-Cre mouse and human pancreatic tumors. Osteopontin/secreted phosphoprotein 1 was identified as the top differentially overexpressed gene in CAF-CM–treated PC cells and knockdown of osteopontin/secreted phosphoprotein 1 significantly reduced stemness characteristics in CAF-CM–treated PC cells. Conclusions: Our data uncovered novel insight into the interplay between CAF and enrichment of stemness population through the osteopontin/secreted phosphoprotein 1–CD44 axis in PC.
KW - CD44
KW - Cancer Stem Cells
KW - Cancer-Associated Fibroblast
KW - OPN/SPP1
KW - Pancreatic Cancer
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U2 - 10.1053/j.gastro.2021.08.023
DO - 10.1053/j.gastro.2021.08.023
M3 - Article
C2 - 34418441
AN - SCOPUS:85116579210
SN - 0016-5085
VL - 161
SP - 1998-2013.e7
JO - Gastroenterology
JF - Gastroenterology
IS - 6
ER -