Integrin α-3 ß-1’s central role in breast cancer, melanoma and glioblastoma cell aggregation revealed by antibodies with blocking activity

Lusche, Daniel F.; Klemme, Michael R.; Soll, David R.; Reis, Ryan; Forrest, Cristopher C.; Nop, Tiffany S.; Wessels, Deborah; Berger, Brian B.; Glover, Rebecca A.

doi:10.1080/19420862.2019.1583987

Cited by 9 publications

(13 citation statements)

References 84 publications

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“…mAb NZ-1, described above, was affinity purified using a Protein G HP spin trap column (GE Healthcare, Cat.#28903134) and concentrated using an ultra-centrifugal filter (EMD Millipore, Cat.# UFC905024) [27]. The concentration was measured by a Nanodrop 1000 spectrophotometer (ThermoScientific) and 0.67 μg added to 4x10 4 F-NHDFs or CC-NHDFs in 200 μl of medium.…”

Section: Methodsmentioning

confidence: 99%

Reciprocal signaling and direct physical interactions between fibroblasts and breast cancer cells in a 3D environment

et al. 2019

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Tumorigenic cells undergo cell aggregation and aggregate coalescence in a 3D Matrigel environment. Here, we expanded this 3D platform to assess the interactions of normal human dermal fibroblasts (NHDFs) and human primary mammary fibroblasts (HPMFs) with breast cancer-derived, tumorigenic cells (MDA-MB-231). Medium conditioned by MDA-MB-231 cells activates both types of fibroblasts, imbuing them with the capacity to accelerate the rate of aggregation and coalescence of MDA-MB-231 cells more than four fold. Acceleration is achieved 1) by direct physical interactions with MDA-MB-231 cells, in which activated fibroblasts penetrate the MDA-MB-231/Matrigel 3D environment and function as supporting scaffolds for MDA-MB-231 aggregation and coalescence, and 2) through the release of soluble accelerating factors, including matrix metalloproteinase (MMPs) and, in the case of activated NHDFs, SDF-1α/CXCL12. Fibroblast activation includes changes in morphology, motility, and gene expression. Podoplanin (PDPN) and fibroblast activation protein (FAP) are upregulated by more than nine-fold in activated NHDFs while activated HPMFs upregulate FAP, vimentin, desmin, platelet derived growth factor receptor A and S100A4. Overexpression of PDPN, but not FAP, in NHDF cells in the absence of MDA-MB-231-conditioned medium, activates NHDFs. These results reveal that complex reciprocal signaling between fibroblasts and cancer cells, coupled with their physical interactions, occurs in a highly coordinated fashion that orchestrates aggregation and coalescence, behaviors specific to cancer cells in a 3D environment. These in vitro interactions may reflect events involved in early tumorigenesis, particularly in cases of field cancerization, and may represent a new mechanism whereby cancer-associated fibroblasts (CAFs) promote tumor growth.

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Section: Methodsmentioning

confidence: 99%

Reciprocal signaling and direct physical interactions between fibroblasts and breast cancer cells in a 3D environment

et al. 2019

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“…Further, pancreatic cancer patients with high expression of Integrin α3 had lower median survival than the comparable low expressing group [58]. Moreover, a recent study with blocking antibodies against integrin β1 or α3 in vitro revealed integrin α3β1 disruption of cell aggregation and aggregate coalescence in breast cancer, melanoma and glioblastoma cells [59]. Thus, CAF-regulated ECM remodeling in the TME enables CAFs and cancer cells to communicate with each other by exchanging signals that are mainly mediated through integrins, consequently supporting cancer progression.…”

Section: Communication Between Cafs and Cancer Cells Mediated By Imentioning

confidence: 99%

Integrins, CAFs and Mechanical Forces in the Progression of Cancer

Jang

Beningo

2019

Cancers

123

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Cells respond to both chemical and mechanical cues present within their microenvironment. Various mechanical signals are detected by and transmitted to the cells through mechanoreceptors. These receptors often contact with the extracellular matrix (ECM), where the external signals are converted into a physiological response. Integrins are well-defined mechanoreceptors that physically connect the actomyosin cytoskeleton to the surrounding matrix and transduce signals. Families of α and β subunits can form a variety of heterodimers that have been implicated in cancer progression and differ among types of cancer. These heterodimers serve as the nexus of communication between the cells and the tumor microenvironment (TME). The TME is dynamic and composed of stromal cells, ECM and associated soluble factors. The most abundant stromal cells within the TME are cancer-associated fibroblasts (CAFs). Accumulating studies implicate CAFs in cancer development and metastasis through their remodeling of the ECM and release of large amounts of ECM proteins and soluble factors. Considering that the communication between cancer cells and CAFs, in large part, takes place through the ECM, the involvement of integrins in the crosstalk is significant. This review discusses the role of integrins, as the primary cell-ECM mechanoreceptors, in cancer progression, highlighting integrin-mediated mechanical communication between cancer cells and CAFs.

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“…mAb concentration and purification were performed as previously described [14,15]. In brief, mAb supernatant was concentrated in VivaCell centrifugal filters with a 50 kDa cut off PLOS ONE (Satorius, Stonehouse, UK).…”

Section: Mab Concentration and Purificationmentioning

confidence: 99%

“…CD44, a transmembrane glycoprotein with a large extracellular domain [1][2][3][4][5], was originally identified as a receptor for the extracellular matrix molecule hyaluronic acid [6][7][8][9]. Subsequently, it was shown that CD44 played a role in a number of cellular processes related to adhesion and cell motility [2,[10][11][12][13], in some cases in the absence of hyaluronic acid [14,15]. CD44 is expressed in a variety of cell types [16][17][18], and has been shown to be up-regulated in stem cells [19][20][21][22] as well as select carcinomas [23][24][25][26][27][28][29][30][31].…”

Section: Introductionmentioning

confidence: 99%

“…Anti-CD44 monoclonal antibodies (mAbs) have been shown to block the formation of subcutaneous tumors and to repress metastasis in mice [32,33], and down regulation of CD44 in cancer cells has been shown to reduce stem cell-associated traits [34][35][36]. Moreover, anti-CD44 mAbs have been shown to affect cancer cell motility and aggregation of breast tumor and melanoma cell lines in a 3D Matrigel model, in the apparent absence of hyaluronic acid (HA) [14,15].…”

Section: Introductionmentioning

confidence: 99%

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New monoclonal antibodies that recognize an unglycosylated, conserved, extracellular region of CD44 in vitro and in vivo, and can block tumorigenesis

et al. 2021

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CD44 is a transmembrane glycoprotein that binds to hyaluronic acid, plays roles in a number of cellular processes and is expressed in a variety of cell types. It is up-regulated in stem cells and cancer. Anti-CD44 monoclonal antibodies affect cell motility and aggregation, and repress tumorigenesis and metastasis. Here we describe four new anti-CD44 monoclonal antibodies originating from B cells of a mouse injected with a plasmid expressing CD44 isoform 12. The four monoclonal antibodies bind to the terminal, extracellular, conserved domain of CD44 isoforms. Based on differences in western blot patterns of cancer cell lysates, the four anti-CD44 mAbs separated into three distinct categories that include P4G9, P3D2, and P3A7, and P3G4. Spot assay analysis with peptides generated inEscherichia colisupport the conclusion that the monoclonal antibodies recognize unglycosylated sequences in the N-terminal conserved region between amino acid 21–220, and analyses with a peptide generated in human embryonic kidney 293 cells, demonstrate that these monoclonal antibodies bind to these peptides only after deglycosylation. Western blots with lysates from three cancer cell lines demonstrate that several CD44 isoforms are unglycosylated in the anti-CD44 target regions. The potential utility of the monoclonal antibodies in blocking tumorigenesis was tested by co-injection of cells of the breast cancer-derived tumorigenic cell line MDA-MB-231 with the anti-CD44 monoclonal antibody P3D2 into the mammary fat pads of mice. All five control mice injected with MDA-MB-231 cells plus anti-IgG formed palpable tumors, while only one of the six test mice injected with MDA-MB-231 cells plus P3D2 formed a tiny tumor, while the remaining five were tumor-free, indicating that the four anti-CD44 mAbs may be useful therapeutically.

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Integrin α-3 ß-1’s central role in breast cancer, melanoma and glioblastoma cell aggregation revealed by antibodies with blocking activity

Cited by 9 publications

References 84 publications

Reciprocal signaling and direct physical interactions between fibroblasts and breast cancer cells in a 3D environment

Reciprocal signaling and direct physical interactions between fibroblasts and breast cancer cells in a 3D environment

Integrins, CAFs and Mechanical Forces in the Progression of Cancer

New monoclonal antibodies that recognize an unglycosylated, conserved, extracellular region of CD44 in vitro and in vivo, and can block tumorigenesis

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