The Untold Story of Granzymes in Oncoimmunology: Novel Opportunities with Old Acquaintances

doi:10.1016/j.trecan.2017.04.001

Trends in Cancer

Volume 3, Issue 6, June 2017, Pages 407-422

https://doi.org/10.1016/j.trecan.2017.04.001 Get rights and content

Trends

GZMs are a family of serine proteases that were originally identified in the cytotoxic granules of cytotoxic T and NK cells. The most-abundant and best-characterized members, GZMA and GZMB, have been found in several immune and, more recently, non-immune cell types.

Several non-cytotoxic functions of GZMs have been demonstrated in mouse models of disease. Some are closely related to processes involved in cancer initiation/progression, including inflammation, angiogenesis, extracellular matrix remodeling, and/or immunosuppression.

Mice deficient in GZMB are protected from the development of some, but not all, types of tumors, and this is related to impaired T regulatory function.

Mice deficient in individual GZMs (especially GZMA) are not compromised in controlling most pathogens and tumors. Thus, inhibition of GZMA should not compromise the protective host immune response.

For more than 20 years perforin and granzymes (GZMs) have been recognized as key cell death executors of cytotoxic T (Tc) and natural killer (NK) cells during cancer immunosurveillance. In immune surveillance, perforin and GZMB, the most potent cytotoxic molecules, act mainly as antitumoral and anti-infectious factors. However, when expressed by immune regulatory cells they may contribute to immune evasion of specific cancer types. By contrast, the other major granzyme, GZMA, seems not to play a major role in Tc/NK cell-mediated cytotoxicity, but acts as a proinflammatory cytokine that might contribute to cancer development. Members of the GZM family also regulate other biological processes unrelated to cell death, such as angiogenesis, vascular integrity, extracellular matrix remodeling, and barrier function, all of which contribute to cancer initiation and progression. Thus, a new paradigm is emerging in the field of oncoimmunology. Can GZMs act as protumoral factors under some circumstances? We review the diverse roles of GZMs in cancer progression, and new therapeutic opportunities emerging from targeting these protumoral roles.

Section snippets

The Changing View of GZMs in Cancer Immunity

The immune system has evolved a vast array of mechanisms to protect the host from pathogens and cancer while maintaining self-tolerance to prevent autoimmune reactions and other inflammatory disorders. Among these mechanisms, natural killer (NK) and cytotoxic T (Tc) cells (including αβ CD8⁺ T, γδ CD8⁺ T, and γδ CD8⁻ T cells) recognize and kill pathogen-infected and cancer cells 1, 2. However, if uncontrolled, Tc and NK cells contribute to the pathology observed in inflammatory and autoimmune

GZMs As Immune Checkpoints

The cellular expression of GZMs is not restricted to Tc and NK cells, or to other populations involved in cancer immunosurveillance (Box 1 and Table 1). Other cells expressing GZMs, such as CD4⁺ T regulatory (Treg) lymphocytes, myeloid-derived suppressor cells (MDSCs), and dendritic cells (DCs), are involved in the homeostatic regulation of proinflammatory T cell activity [i.e., type 1 T helper cell (Th1), Th17, or Tc subsets], preventing autoimmune and inflammatory reactions. These types of

GZMs and Inflammation

GZMs are emerging also as key regulators of inflammation-related disorders (Box 2), and could thus contribute to cancer initiation and progression, as shown in inflammatory cancers [34]. Although several GZMs, including human and mouse GZMA, GZMM, GZMB, or GZMK, are able to modulate proinflammatory responses in different cell types (monocytes, macrophages, endothelial, epithelial, and fibroblast cells) in vitro 35, 36, 37, 38, 39, 40, 41, 42 and in vivo 31, 36, 43, 44, the available evidence

GZMs, ECM Remodeling, and Cancer Progression: A Time for Speculation

The ECM is a key component of the tumor microenvironment and a dynamic structure that regulates several steps of the tumor life cycle. ECM regulation of angiogenesis and receptor signaling are of special interest in cancer biology because they are involved in survival, proliferation, invasion, and metastasis.

The tumor microenvironment is rich in serine proteases that are usually produced by stromal and cancer cells, and which cleave ECM proteins that modulate angiogenesis and metastasis. Among

Other Roles of GZMs in Cancer Progression

Recent work has suggested a protumoral role of GZMM in EMT [66]. GZMM expression in human cancerous tissue correlates with cells expressing an EMT phenotype, and cancer cell lines in which GZMM was downregulated by shRNA induced less metastasis in mouse models. The mechanisms underlying this process are not clear. Although the authors suggested that GZMM activates STAT3 signaling, it is still not clear how this pathway could be activated in a culture of CRC lines where cytokines such as IL-6

GZMs As Therapeutic Targets in Cancer – Can Cytotoxic Effects Be Reduced?

The accumulated knowledge of immunoregulatory cell networks in cancer development and progression has allowed the development of more effective immunotherapies. We now know the key molecules to target to reduce chronic inflammation and thus prevent cancer onset and/or progression (i.e., UC-CRC or asbestos in mesothelioma) 3, 34, and more specific anti-inflammatory agents have been developed such as TNF-α or IL-6 blockers 68, 69. In addition, antibodies that block inhibitory immune checkpoints

Concluding Remarks

Accumulating evidence in mouse and human models suggests that GZMs are crucial regulators of many processes unrelated to cell death, such as inflammation, angiogenesis, ECM remodeling, and immune homeostasis. These novel functions of GZMs challenge the traditional dogma of a protective role of these proteases in cancer immunosurveillance and immunotherapy. Indeed, many processes regulated by GZMs are key promoters of cancer initiation and progression. The balance between antitumoral and

Acknowledgments

This work was supported in part by Fondo Social Europeo (J.P.) and Ministerio de Economia y Competitividad (SAF2011-25390) (J.P.) and SAF2014-54763-C2-1 (J.P.). L.S. and M.A. were supported by a grant from Fundación Santander/Universidad de Zaragoza and FPI/Ministerio de Economia y Competitividad (L.S.). P.J.S. was supported by a predoctoral contract from Gobierno de Aragon. J.P. was supported by Fundación Aragon I+D (ARAID). D.J.G. is funded by a Canadian Institutes for Health Research (CIHR)

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Citation Excerpt :
GZMA, which is also called granzyme A, is a dimer with tryptase activity and acts on the level of the cell nucleus, which leads to DNA damage. Recent evidence has shown that GZMA does not play a major role in Tc/NK cell-mediated cytotoxicity, but acts as a proinflammatory cytokine that might contribute to cancer development [49]. In terms of IFN-γ (interferon-γ), it normally plays an important role in anti-tumor immunity.
Interleukin-1α (IL-1α) is thought to play a major role in renal cell carcinoma (RCC), but the specific mechanism is unclear. In this study, we explored the potential mechanism of the role of IL-1α in RCC.
Two cohorts were included in this study: The Cancer Genome Atlas (TCGA) cohort (n=521) and Fudan University Shanghai Cancer Center (FUSCC) cohort (n=198). These cohorts were used for cell infiltration-related analyses.
In TCGA cohort, expression of IL-1α was significantly increased in patients with RCC. High IL-1α mRNA expression and intratumoral IL-1α+ cells were correlated with poor OS, which remained significant after adjustment for confounders. Elevated IL-1α mRNA expression and increased intratumoral IL-1α+ cell infiltration were associated with less mast cell infiltration in RCC. Among the relationships between IL-1α and cytotoxic factors, IL-1α was correlated with IFN-γ, TNFSF11, and GZMA. Among the relationships between IL-1α and immunosuppressive factors, we found that IL-1α was correlated with PDCD1, CD274, CTL1A4, LAG3, and BTLA.
Expression of IL-1α was significantly correlated with the prognosis of RCC patients. IL-1α may participate in the development and progression of renal cancer through interactions with immune infiltrating mast cells. Our data demonstrate that IL-1α is a prognostic factor and potential therapeutic target for RCC.
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GZMK can serve as an accelerator of inflammatory processes that directly induce proinflammatory cytokine release, reactive oxygen species generation, endothelial dysfunction and subsequent immune cell recruitment [14,15]. As a secreted molecule, GZMK can also be found in the extracellular milieu where they will exert similar regulating functions [16,17]. In healthy individuals, serum GZMK levels were low or undetectable, whereas levels of soluble GZMK were notably elevated in the circulation under some inflammatory conditions [18–20].
Immune inflammatory dysfunction is a hallmark of abdominal aortic aneurysm (AAA). Granzyme K (GZMK) is involved in the regulation of inflammation. However, the correlation between GZMK expression and AAA risk remains unknown.
This case-control study included 112 AAA patients and 112 controls. Serum GZMK levels were determined by enzyme-linked immunosorbent assay and immunohistochemistry was utilized to determine GZMK expression in aortic tissues.
Compared with controls, AAA patients had higher levels of serum GZMK, and GZMK expression in AAA tissues was increased and positively associated with its serum levels (r = 0.688, P = 0.019). A positive association of serum GZMK levels with CRP or AAA diameter was confirmed, while there was a relationship between tissue GZMK expression and AAA diameter. The AUC of serum GZMK for AAA diagnosis was 0.78 with the sensitivity and specificity of 62.5% and 81.2%, whereas AUC for rupture detection was 0.76 with a sensitivity of 90.0% and specificity of 51.3%. A combination of clinically used inflammatory parameters with serum GZMK could enhance the accuracy of WBC or CRP alone in detecting AAA or rupture type. Multiple logistic analyses revealed an association of per unit increase of serum GZMK with AAA presence (OR = 1.046, P < 0.001) and its rupture risk (OR = 1.015, P = 0.048) after adjusting for confounding factors.
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Trends in Cancer

ReviewThe Untold Story of Granzymes in Oncoimmunology: Novel Opportunities with Old Acquaintances

Trends

Section snippets

The Changing View of GZMs in Cancer Immunity

GZMs As Immune Checkpoints

GZMs and Inflammation

GZMs, ECM Remodeling, and Cancer Progression: A Time for Speculation

Other Roles of GZMs in Cancer Progression

GZMs As Therapeutic Targets in Cancer – Can Cytotoxic Effects Be Reduced?

Concluding Remarks

Acknowledgments

Trends Cancer

Blood

Immunity

Immunity

Gastroenterology

Blood

Blood

Mol. Immunol.

Mol. Immunol.

Lab. Invest.

Am. J. Pathol.

J. Biol. Chem.

Cell Immunol.

Immunity

Mol. Cell

Cell Rep.

Immunity

Trends Cancer

J. Biol. Chem.

Blood

Cancer Cell

Am. J. Transpl.

Trends Immunol.

Immunobiology

Immunol. Lett.

How do cytotoxic lymphocytes kill cancer cells?

Clin. Cancer Res.

Natural innate and adaptive immunity to cancer

Annu. Rev. Immunol.

Cancer-related inflammation

Nature

Positive and negative regulation of cellular immune responses in physiologic conditions and diseases

Clin. Dev. Immunol.

Mucosal immunity to pathogenic intestinal bacteria

Nat. Rev. Immunol.

The tumor microenvironment at a glance

J. Cell Sci.

Contact-mediated suppression by CD4+CD25+ regulatory cells involves a granzyme B-dependent, perforin-independent mechanism

J. Immunol.

Low expression of Granzyme B in colorectal cancer is associated with signs of early metastastic invasion

Histopathology

Effector memory T cells, early metastasis, and survival in colorectal cancer

N. Engl. J. Med.

The ratio of FOXP3+ regulatory T cells to granzyme B+ cytotoxic T/NK cells predicts prognosis in classical Hodgkin lymphoma and is independent of Bcl-2 and MAL expression

Am. J. Clin. Pathol.

Foxp3+ cell infiltration and granzyme B+/Foxp3+ cell ratio are associated with outcome in neoadjuvant chemotherapy-treated ovarian carcinoma

Cancer Immunol. Immunother.

Absence of granzyme B positive tumour-infiltrating lymphocytes in primary melanoma excisional biopsies is strongly associated with the presence of sentinel lymph node metastasis

Cell Oncol.

FOXP3 defines regulatory T cells in human tumor and autoimmune disease

Cancer Res.

Interleukin 21-induced granzyme B-expressing B cells infiltrate tumors and regulate T cells

Cancer Res.

Granzyme B-mediated degradation of T-cell receptor zeta chain

Cancer Res.

Delivery and therapeutic potential of human granzyme B

Immunol. Rev.

Human B cells differentiate into granzyme B-secreting cytotoxic B lymphocytes upon incomplete T-cell help

Immunol. Cell Biol.

Activated mouse B cells lack expression of granzyme B

J. Immunol.

Induction of myelodysplasia by myeloid-derived suppressor cells

J. Clin. Invest.

Granzyme B is expressed in mouse mast cells in vivo and in vitro and causes delayed cell death independent of perforin

Cell Death Differ.

Control of granzymes by serpins

Cell Death Differ.

Immunity, inflammation, and cancer: an eternal fight between good and evil

Review
The Untold Story of Granzymes in Oncoimmunology: Novel Opportunities with Old Acquaintances

Contact-mediated suppression by CD4⁺CD25⁺ regulatory cells involves a granzyme B-dependent, perforin-independent mechanism

The ratio of FOXP3⁺ regulatory T cells to granzyme B⁺ cytotoxic T/NK cells predicts prognosis in classical Hodgkin lymphoma and is independent of Bcl-2 and MAL expression

Foxp3⁺ cell infiltration and granzyme B⁺/Foxp3⁺ cell ratio are associated with outcome in neoadjuvant chemotherapy-treated ovarian carcinoma