Granzyme B Degraded Type IV Collagen Products in Serum Identify Melanoma Patients Responding to Immune Checkpoint Blockade

Abstract

A T-cell permissive tumor microenvironment, characterized by the presence of activated T cells and low fibrotic activity is crucial for response to immune checkpoint inhibitors (ICIs). Granzyme B has been shown to promote T-cell migration through the basement membrane by the degradation of type IV collagen. In this study, we evaluated the biomarker potential of measuring granzyme B-mediated degradation of type IV collagen (C4G) in combination with a fibroblast activation biomarker (PRO-C3) non-invasively for identifying metastatic melanoma patients responding to the ICI ipilimumab. A monoclonal antibody was generated against C4G and used to develop a competitive electro-chemiluminescence immunoassay. C4G and PRO-C3 were measured in pretreatment serum from metastatic melanoma patients (n = 54). The C4G assay was found specific for a granzyme B-generated neo-epitope on type IV collagen. The objective response rate (ORR) was 2.6-fold higher (18% vs. 7%) in patients with high C4G levels (>25th percentile) vs. low levels (≤25th percentile). Likewise, high C4G levels at baseline were associated with longer overall survival (OS) (log-rank, p = 0.040, and hazard ratio (HR) = 0.48, 95%CI: 0.24–0.98, p = 0.045). Combining high C4G with low PRO-C3 correlated with improved OS with a median OS of 796 days vs. 273 days (p = 0.0003) and an HR of 0.30 (95%CI: 0.15–0.60, p = 0.0006). In conclusion, these results suggest that high granzyme B degraded type IV collagen (C4G) combined with low PRO-C3 quantified non-invasively has the potential to identify the responders to ICI therapy.

Cancer immunotherapy by NC410, a LAIR-2 Fc protein blocking human LAIR-collagen interaction

“…During ECM turnover, proteolytically cleaved matrix degradation fragments are released into the systemic circulation (Bonnans, Chou et al 2014).Serum levels of collagen degradation fragments are elevated in cancer patients compared to healthy controls, and checkpoint blockade alters serum CDP levels (Jensen, Madsen et al 2018, Jensen, Sinkeviciute et al 2020, Wang, Bager et al 2021). NC410 treatment increased specific fragments generated by degradation of collagen VI by MMP-2 (C6M) and collagen IV by granzyme B (C4G).…”

“…We therefore hypothesize that specific granzyme B degraded type IV collagen fragments and other fragments are released to the circulation when T cells infiltrate tumors and may have potential as a clinical pharmacodynamic biomarker. Interestingly, granzyme B degraded type IV collagen fragments (C4G) associate with favorable anti-CTLA-4 treatment response in metastatic melanoma patients (Jensen, Madsen et al 2018, Jensen, Sinkeviciute et al 2020. (St-Pierre and Potworowski 2000)The potential of C6M and C4G collagen fragments as biomarkers of clinical response will be further investigated in a recently initiated NC410 first-in-human clinical trial (NCT04408599).…”

“…Recently it has been proposed that specific collagen degradation products (CDPs) reflecting the turnover of specific collagens may be used as biomarkers to non-invasively (in serum) interrogate cell reactivity in the TME and predict response to treatment (Jensen, Madsen et al 2018, Jensen, Sinkeviciute et al 2020, Wang, Bager et al 2021. Because NC410 both engages collagens and promotes local T cell responses, it was surmised that NC410 treatment may result in changes in specific CDP levels, which potentially be indicative of tumor collagen remodeling.…”

Noninvasive prognostic biomarker potential of quantifying the propeptides of Type XI collagen alpha‐1 chain (PRO‐C11) in patients with pancreatic ductal adenocarcinoma

“…In addition, our group has recently showed the importance of targeting specific PTMs of Type IV collagen and vimentin in metastatic melanoma and lung cancer. We demonstrated that two different neo‐epitopes on the same protein (Type IV collagen and vimentin) could in fact provide opposite prognostic value 37,38 . These data underline the importance of targeting the relevant epitope in cancer biomarker discovery.…”

“…We demonstrated that two different neo-epitopes on the same protein (Type IV collagen and vimentin) could in fact provide opposite prognostic value. 37,38 These data underline the importance of targeting the relevant epitope in cancer biomarker discovery. Furthermore, it urges for caution when targeting the full-length protein, instead of targeting protein fragments as also suggested by Hosein et al 6 In our study, we observed that targeting two different neo-epitopes located in the propeptide of Type XI collagen had completely differential biomarker value.…”