Low molecular weight hyaluronan inhibits colorectal carcinoma growth by decreasing tumor cell proliferation and stimulating immune response
Introduction
Colorectal carcinoma is the second most common cancer worldwide [1], [2] and surgery remains the primary treatment. However it can only be performed in 20% of patients with metastatic disease, with a 5-year survival rate of 25–40%, despite adjuvant chemotherapy [3]. Therefore, new therapeutic options are urgently needed for the advanced disease. At the moment, most of research is focused on the development of new therapeutic strategies, such as blockage/inhibition of signal transduction mechanisms (i.e. targeting the epidermal growth factor receptor or Ras), sequestering vascular endothelial growth factor receptor ligands and/or enhancing immune response mechanisms [4], [5]. Immunotherapeutic strategies hold promise; however, they still remain as an experimental discipline. Despite that CRC has been considered poorly immunogenic, it has been found that the presence of lymphocyte infiltration in tumors is correlated with patient survival, showing higher survival rate in comparison with those with low density of T-cell infiltrates [6]. For this reason it is necessary to find an immune adjuvant or to artificially up-regulate the immune response in order to allow a curative effect.
Hyaluronan (HA) is a lineal, large and ubiquitous glycosaminoglycan found in almost all tissues and especially in those undergoing cell proliferation, regeneration and repair, such as embryonic, inflammated and tumor stroma tissues [7]. It is well established that HA functions are size-dependent: in normal tissues HMW HA (≈106 Da) is the predominant form and has a homeostatic role in regulating cell behavior. During the repair, inflammation and tumor processes, there is a predominance of LMW HA fragments (≈105 Da) which have been shown to promote cell proliferation, adhesion and motility [8]. However, smaller HA fragments (≈2.5 × 103) were found to disrupt HA-CD44 interaction and to induce tumor cell death [7]. The effects of HA and its fragments have been shown to be mediated by the PI3K/Akt/NFkB pathway in different tumors such as breast cancer, melanoma, colorectal carcinoma and lymphoma [9], [10], [11].
During the inflammation process, HMW HA can be depolymerized to LMW fragments via oxygen radicals and enzymatic degradation by hyaluronidase, glucuronidase, and hexosaminidase [12]. LMW HA form has been shown to up-regulate the expression of inflammatory genes in epithelial, endothelial and DCs as well as in fibroblasts and macrophages. Genes induced by LMW HA include members of the chemokine family, cytokines (IL-8, IL-12 and TNF-α) and inducible NO synthase [13], [14]. In addition, LMW HA or its fragments were shown to enhance T cell responses by activating and up-regulating costimulatory molecules on DCs [15], [16]. On the contrary, recent evidence suggests that HMW HA has an immunosuppressive role instead [17], [18].
Several studies have shown association of HA biosynthesis alteration with cancer progression (including CRC), as well as an increase in HA serum levels and tumor stroma deposition [19], [20], [21], [22]. It is assumed that the predominance of LMW HA in tumors is a result of tissue injury and that it is caused by a lack of balance in between HA synthases (Has) and hyaluronidases (Hyal) activities. The LMW HA found within tumor microenvironment is considered to be a tumor growth promoter [8], [23], [24]. Nonetheless, few studies have shown the exact molecular weight of HA in tumors and its effect on the antitumoral immune response [15], [19]. Therefore, assessment of the exact HA molecular weight form and its role in immune regulation during cancer progression remains largely unexplored.
In this work we have addressed the in vivo effects of exogenous application of both LMW- and HMW-HA into a murine model of CRC.
Section snippets
Reagents
HA recombinant of definite size (HMW 1.5–1.8 × 106 Da; LMW 1–3 × 105 Da; from CPN spol.s.r.o. Czech Republic) was kindly supplied by Farmatrade (Bs. As., Argentina). Oligosaccharides were obtained by enzymatic digestion from HMW-HA, as previously described [25]. This method render fragments ranging from HA4 tetrasaccharides (two disaccharide units in length: 700 Da) to HA14 oligosaccharides (seven disaccharide units in length: 2600 Da). HA-FITC from bovine trachea (≈1 × 105 Da) was purchased to Calbiochem.
Results and discussion
It has been previously described that HA modulate the behavior of different tumors by multiple mechanisms [7], [19], [31]. These effects are dependent on HA molecular weight, concentration and interaction with other extracellular matrix components present in tumor microenvironment [7], [8], [31]. Thus, we decided to examine in vitro and in vivo the pro- or anti tumoral effect of different types of HA in an experimental CRC model. In order to analyze if HA could directly modify CT26 behavior, we
Conflict of interest statement
All the authors declare that they do not have conflict of interest to disclose and they did not receive honoraria and support from any pharmaceutical company.
Acknowledgments
This work was supported by grants from Mizutani Foundation for Glycoscience (80072), Universidad Austral and Agencia Nacional de Promoción Científica y Tecnológica (PICT-2006-1882; PICT-2005-34788, PICTO-CRUP 2005-31179), CTE-06 and AECI 2006. We acknowledge the continuous support from Inés Bemberg. We thank Soledad Arregui and Guillermo Gastón for expert technical assistance.
References (37)
- et al.
Colorectal cancer
Lancet
(2005) - et al.
Therapeutic vaccination in patients with gastrointestinal malignancies. A review of immunological and clinical results
Ann. Oncol.
(2005) - et al.
Hyaluronan oligosaccharides inhibit anchorage-independent growth of tumor cells by suppressing the phosphoinositide 3-kinase/Akt cell survival pathway
J. Biol. Chem.
(2002) - et al.
Hyaluronan-mediated CD44 interaction with RhoGEF and Rho kinase promotes Grb2-associated binder-1 phosphorylation and phosphatidylinositol 3-kinase signaling leading to cytokine (macrophage-colony stimulating factor) production and breast tumor progression
J. Biol. Chem.
(2003) - et al.
Hyaluronan fragments induce nitric-oxide synthase in murine macrophages through a nuclear factor kappa B-dependent mechanism
J. Biol. Chem
(1997) - et al.
Hyaluronan – magic glue for the regulation of the immune response?
Trends Immunol.
(2003) - et al.
Altered hyaluronan biosynthesis in cancer progression
Semin. Cancer Biol.
(2008) - et al.
Hyaluronan constitutively regulates activation of COX-2-mediated cell survival activity in intestinal epithelial and colon carcinoma cells
J. Biol Chem.
(2008) Hyaluronan and its catabolic products in tissue injury and repair
Matrix Biol.
(2002)- et al.
Hyaluronan oligosaccharides inhibit tumorigenicity of osteosarcoma cell lines MG-63 and LM-8 in vitro and in vivo via perturbation of hyaluronan-rich pericellular matrix of the cells
Am. J. Pathol.
(2007)
Global Cancer Statistics, 2002, CA
Cancer J. Clin.
Systemic treatment of colorectal cancer
Gastroenterology
Immunotherapy and immunoescape in colorectal cancer
World J. Gastroenterol.
Type, density, and location of immune cells within human colorectal tumors predict clinical outcome
Science
Hyaluronan: from extracellular glue to pericellular cue
Nat. Rev. Cancer
Hyaluronan in tissue injury and repair
Ann. Rev. Cell Dev. Biol.
Hyaluronan oligosaccharides sensitize lymphoma resistant cell lines to vincristine by modulating P-glycoprotein activity and PI3K/Akt pathway
Int. J. Cancer
Hyaluronan: its nature, distribution, functions and turnover
J. Intern. Med.
Cited by (59)
Marine polysaccharides: Biological activities and applications in drug delivery systems
2024, Carbohydrate ResearchExploiting ECM remodelling to promote immune-mediated tumour destruction
2022, Current Opinion in ImmunologyCitation Excerpt :Alaniz and colleagues found that LMW-HA treated mice exhibited increased expression of MHC-II and CD80 on DCs, favouring antigen presentation in secondary lymphoid organs. This was associated with enhanced T cell infiltration in the tumour and significantly reduced tumour growth [37]. A subsequent study conducted on DCs isolated from colorectal cancer showed that after treatment with LMW-HA, CCR7 expression on DCs as well as MMP activity was increased.
Expression and characterization of a thermotolerant and pH-stable hyaluronate lyase from Thermasporomyces composti DSM22891
2021, Protein Expression and PurificationCellular hyaluronan is associated with a poor prognosis in renal cell carcinoma
2020, Urologic Oncology: Seminars and Original InvestigationsControlled conversion of sodium hyaluronate into low-molecular-weight polymers without additives using high-temperature water and fast-heating-rates
2020, Journal of Supercritical FluidsSynthesis and study of branched hyaluronic acid with potential anticancer activity
2019, Carbohydrate Polymers