TY - JOUR
T1 - Ablation of VLA4 in multiple myeloma cells redirects tumor spread and prolongs survival
AU - Hathi, Deep
AU - Chanswangphuwana, Chantiya
AU - Cho, Nicholas
AU - Fontana, Francesca
AU - Maji, Dolonchampa
AU - Ritchey, Julie
AU - O’Neal, Julie
AU - Ghai, Anchal
AU - Duncan, Kathleen
AU - Akers, Walter J.
AU - Fiala, Mark
AU - Vij, Ravi
AU - DiPersio, John F.
AU - Rettig, Michael
AU - Shokeen, Monica
N1 - Funding Information:
Optical imaging (IVIS Spectrum CT) was supported by NIH -S10OD027042. This research was also supported by the Alvin J. Siteman Cancer Center through The Foundation for Barnes-Jewish Hospital and the National Cancer Institute (P30 CA091842). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. We thank the patients who provided samples to the Multiple Myeloma Tissue Bank Study at Washington University that were used in this study. We thank Dr. Shondra Miller and the Genome Engineering & iPSC Center for their wonderful assistance with the generation and analysis of the 5TGM1 ITGA4 KO clones.
Funding Information:
We thank Professor Samuel Achilefu for his insights. Gail Sudlow assisted in intravenous (i.v.) tail vein injections of 5TGM1-GFP cells and LLP2A-Cy5 in C57Bl/KaLwRij mice. We thank Noriko Yanaba in the laboratory of Professor Gregory Lanza for help with preparation of histology slides. This work has been supported by R01 CA248493 [Shokeen], the NCI funded Center for Multiple Myeloma Nanotherapy [U54 CA199092], the Imaging Sciences Pathway (ISP) fellowship [T32 EB014855], Molecular Imaging Center [NIH P50 CA094056], Siteman Cancer Center Small Animal Cancer Imaging shared resource [NCI P30 CA091842], and the Nutrition Obesity Research Center [P30 DK056341]. Live cell fluorescence microscopy was performed in part through the use of Washington University Center for Cellular Imaging (WUCCI) supported by Washington University School of Medicine, The Children’s Discovery Institute of Washington University and St. Louis Children’s Hospital (CDI-CORE-2015-505 and CDI-CORE-2019-813) and the Foundation for Barnes-Jewish Hospital (3770 and 4642).
Publisher Copyright:
© 2022, The Author(s).
PY - 2022/12
Y1 - 2022/12
N2 - Multiple myeloma (MM) is a cancer of bone marrow (BM) plasma cells, which is increasingly treatable but still incurable. In 90% of MM patients, severe osteolysis results from pathological interactions between MM cells and the bone microenvironment. Delineating specific molecules and pathways for their role in cancer supportive interactions in the BM is vital for developing new therapies. Very Late Antigen 4 (VLA4, integrin α4β1) is a key player in cell–cell adhesion and signaling between MM and BM cells. We evaluated a VLA4 selective near infrared fluorescent probe, LLP2A-Cy5, for in vitro and in vivo optical imaging of VLA4. Furthermore, two VLA4-null murine 5TGM1 MM cell (KO) clones were generated by CRISPR/Cas9 knockout of the Itga4 (α4) subunit, which induced significant alterations in the transcriptome. In contrast to the VLA4+ 5TGM1 parental cells, C57Bl/KaLwRij immunocompetent syngeneic mice inoculated with the VLA4-null clones showed prolonged survival, reduced medullary disease, and increased extramedullary disease burden. The KO tumor foci showed significantly reduced uptake of LLP2A-Cy5, confirming in vivo specificity of this imaging agent. This work provides new insights into the pathogenic role of VLA4 in MM, and evaluates an optical tool to measure its expression in preclinical models.
AB - Multiple myeloma (MM) is a cancer of bone marrow (BM) plasma cells, which is increasingly treatable but still incurable. In 90% of MM patients, severe osteolysis results from pathological interactions between MM cells and the bone microenvironment. Delineating specific molecules and pathways for their role in cancer supportive interactions in the BM is vital for developing new therapies. Very Late Antigen 4 (VLA4, integrin α4β1) is a key player in cell–cell adhesion and signaling between MM and BM cells. We evaluated a VLA4 selective near infrared fluorescent probe, LLP2A-Cy5, for in vitro and in vivo optical imaging of VLA4. Furthermore, two VLA4-null murine 5TGM1 MM cell (KO) clones were generated by CRISPR/Cas9 knockout of the Itga4 (α4) subunit, which induced significant alterations in the transcriptome. In contrast to the VLA4+ 5TGM1 parental cells, C57Bl/KaLwRij immunocompetent syngeneic mice inoculated with the VLA4-null clones showed prolonged survival, reduced medullary disease, and increased extramedullary disease burden. The KO tumor foci showed significantly reduced uptake of LLP2A-Cy5, confirming in vivo specificity of this imaging agent. This work provides new insights into the pathogenic role of VLA4 in MM, and evaluates an optical tool to measure its expression in preclinical models.
UR - http://www.scopus.com/inward/record.url?scp=85122484256&partnerID=8YFLogxK
U2 - 10.1038/s41598-021-03748-0
DO - 10.1038/s41598-021-03748-0
M3 - Article
C2 - 34996933
AN - SCOPUS:85122484256
SN - 2045-2322
VL - 12
JO - Scientific Reports
JF - Scientific Reports
IS - 1
M1 - 30
ER -