Collagen-rich Tumors Research Articles

Collagen-disruptive cell therapy: adoptive transfer of membrane-anchored, tumor cell surface vimentin-targeted interleukin 12-armed TILs suppress collagen expression to boost deep T-cell infiltration via dual signaling activation and significant CCKAR reduction.

Tumor-targeted T-cell therapies of various types have been booming, but T-cell therapy is limited by its inability to penetrate the collagen barrier surrounding tumors. The destruction of tumor collagen is significant because collagen both suppresses T cells and contributes to the formation of the extracellular matrix. Our previously reported cell surface vimentin (CSV)-targeted and membrane-anchored IL12-armed (attIL12) T cells can reduce collagen production by killing cancer-associated fibroblasts, thereby increasing T-cell infiltration. However, attIL12-T cells cannot reduce collagen expression by tumors that highly express CCKAR. In this study, we discovered that CCKAR directly boosts collagen production by tumor cells in vitro and in vivo. attIL12-modified tumor-infiltrating lymphocytes (TILs) disabled collagen production by CCKAR-high autologous tumor cells in vitro and sarcoma patient-derived xenografts (PDXs) in vivo. This disruption of collagen production by tumor cells required a simultaneous interaction between the CSV on autologous tumor cells, which is targeted by attIL12, and HLA-TCR on attIL12-TILs; when either interaction was abrogated, collagen production and CCKAR expression were not shut down. Mechanistically, the interaction between attIL12-TILs and autologous tumor cells synergized IFNγ production, which in combination with CCKAR downregulation reduced collagen expression through suppression of both TGFβ-stimulated SMAD activation and CCKAR-AKT signaling. Diminishing collagen expression from tumor cells significantly increased T-cell infiltration and improved tumor growth inhibition in PDX sarcomas. This study thus uncovers the first tumor collagen-disrupting T-cell therapy we know of. This is significant because collagen is enriched in most high-grade CCKAR+ human sarcomas. Thus, this attIL12-TIL therapy holds great clinical potential for boosting T-cell infiltration in high-grade, collagen-rich tumors.

Abstract 1341: Non-invasive biomarkers quantifying collagens essential for embryogenesis are elevated in the circulation of solid tumor patients and may guide drug development

Abstract Background Collagen remodeling is an essential part of embryogenesis and defines tissue structure and function in adulthood. The 28 different collagens are all important for maintaining tissue homeostasis and are deregulated in cancer. The abundant, major collagens, such as type I collagen, serve a structural role, whereas the less abundant, minor collagens serve a more regulatory role. Fibroblasts are the primary source of collagens and different cancer-associated fibroblast (CAF) subtypes, such as myCAFs or iCAFs, have been attributed important roles in cancer and are the target of novel therapies. Emerging evidence suggests minor collagens may be aberrantly expressed in cancer and linked to specific fibroblast subtypes, suggesting novel biomarker potential. This study aimed to evaluate the association between different CAF subtypes and collagen expression. Methods We investigated collagen expression in pancreatic cells and CAF subtypes in a publicly available single-cell RNA-Seq dataset (GSA: CRA001160). 24 samples were from patients with pancreatic ductal adenocarcinoma (PDAC) as this is one of the most collagen rich tumor types, and 11 samples were from pancreatic cysts, bile duct- or duodenal-tumors. In addition, we generated a collagen turnover profile using a panel of immunoassays to measure specific collagen fragments in the serum of 220 patients with various solid tumor types and 33 healthy controls. The included immunoassays measured collagen epitopes associated with either formation, degradation, or other modifications of collagens. Results Based on the single-cell RNA-Seq dataset, fibroblasts were the primary source of collagens. Major collagens, including COL1A1, COL3A1, COL4A1, and COL6A1, were highly expressed in both healthy fibroblasts and PDAC CAFs, whereas minor collagens such as COL8A1, COL10A1, COL11A1, and COL12A1 were exclusively expressed in CAFs. When further examining the CAF subtypes, major collagens were expressed in all CAF subtypes, with high expression in myCAFs and iCAFs. In contrast, the minor collagens were exclusively expressed in the myCAFs. When measuring these collagens in serum, the minor collagens were elevated across all cancer types when compared to healthy control serum. Contrastingly, the major collagens were not elevated in any cancers or only elevated in a few cancer types. Conclusions Collagens in general are highly expressed in cancer. However, minor collagens may be more cancer-specific than major collagens. Our findings also suggest that minor collagens may be specific to certain subtypes of CAFs. These collagens can be quantified in blood, making them promising actionable biomarkers that may guide drug development aimed at targeting CAFs. Citation Format: Jeppe Thorlacius-Ussing, Christina Jensen, Emilie Madsen, Morten Karsdal, Nicholas Willumsen. Non-invasive biomarkers quantifying collagens essential for embryogenesis are elevated in the circulation of solid tumor patients and may guide drug development [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1341.

Abstract LB219: Preclinical development of NGM438, a novel anti-LAIR1 antagonist monoclonal antibody for the treatment of collagen-rich solid tumors

Abstract Background: Collagen is an abundant component of the extracellular matrix-enriched stromal microenvironment of solid cancers. Emerging research suggests that effective anti-tumor immunity and responsiveness to immune checkpoint inhibitor (CPI) therapy, may be subverted by the presence of collagen and tumor stroma. LAIR1 functions as an inhibitory receptor for collagen, and is highly expressed on tumor-associated macrophages, and to an extent on T cells. LAIR1 expression correlates with poor overall survival and CPI therapy resistance. To test the hypothesis that LAIR1 mediates tumor stroma-driven immune suppression and promotes CPI resistance, we generated NGM438, a monoclonal mAb to antagonize LAIR1 activity in solid cancers. Herein, we describe NGM438, characterize LAIR1 expression and distribution in human tumors, and provide evidence that LAIR1 antagonism reverses collagen-driven immune suppression and improves CPI responsiveness in immune cell-based assays and preclinical animal models. Methods: NGM438 was identified and developed based on its ability to bind specifically to LAIR1, antagonize ligand binding, and reverse collagen-based immune suppression. NGM438 was tested using several immune cell-based functional assays, alone and in combination with the anti-PD1 mAb, pembrolizumab. Expression and distribution of LAIR1 was evaluated using flow cytometry and immunohistochemistry on healthy donor and cancer patient samples. A surrogate anti-mouse LAIR1 antagonist mAb was also developed and evaluated in preclinical animal models in combination with anti-PD1 mAbs. Results: NGM438 reversed collagen-induced immune suppression in myeloid cells and promoted anti-PD1 mAb-driven T cell responses in immune cell-based functional assays. LAIR1 was expressed on circulating immune cells from cancer patients actively receiving CPI therapy, and was elevated on patient-matched intratumoral macrophages. Histological assessments revealed LAIR1 expression on mononuclear immune cells that infiltrated the parenchyma and collagen-rich tumor stroma across indications. Finally, LAIR1 antagonism, in combination with anti-PD1 mAb treatment, led to significant tumor growth inhibition in a preclinical model resistant to either treatment alone. Conclusions: NGM438 is a novel LAIR1 antagonist mAb that reverses collagen-based immune suppression. LAIR1 was expressed on cancer patient circulating and intratumoral immune cells, and LAIR1-expressing cells were often found in collagen-rich tumor stroma. Preclinical data demonstrated that LAIR1 antagonism sensitized a resistant mouse tumor model to respond to anti-PD1 mAb treatment. These data support clinical evaluation of LAIR1 antagonist mAb NGM438. We aim to test the hypothesis that LAIR1 mediates collagen-driven immune suppression, alone and in combination with PD1 inhibition, in patients with solid cancers. As such, we will be advancing NGM438 to the clinic in early 2022. Citation Format: Sisi He, Jiawei Huang, Leticia Rodriguez, Czrina Cortez, Betty Li, Carmence Ho, Amir Ashique, Kalyani Mondal, Vicky Lin, Julie Roda, Hui Tian, Yan Wang, Bin Fan, Igor Mikaelian, James Sissons, Lee Rivera, Don Gibbons, Jonathan Sitrin. Preclinical development of NGM438, a novel anti-LAIR1 antagonist monoclonal antibody for the treatment of collagen-rich solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr LB219.

Impact of Collagen Triple Helix Structure on Melanoma Cell Invadopodia Formation and Matrix Degradation upon BRAF Inhibitor Treatment.

A collagen-rich tumor microenvironment (TME) is associated with worse outcomes in cancer patients and contributes to drug resistance in many cancer types. In melanoma, stiff and fibrillar collagen-abundant tissue is observed after failure of therapeutic treatments with BRAF inhibitors. Increased collagen in the TME can affect properties of the extracellular matrix (ECM), including stiffness, adhesiveness, and interaction of integrins with triple helix forming nanostructures. Decoupling these biochemical and biophysical properties of the ECM can lead to a better understanding of how each of these individual properties affect melanoma cancer behavior and drug efficacy. In addition, as drug treatment can induce cancer cell phenotypic switch, cancer cell responsiveness to the TME can be dynamically changed during therapeutic treatments. To investigate cancer cell phenotype changes and the role of the cancer TME, poly(ethylene glycol) (PEG) hydrogels functionalized with collagen mimetic peptides (CMPs) is utilized, or an interpenetrating network (IPN) of type І collagen within the PEG system to culture various melanoma cell lines in the presence or absence of Vemurafenib (PLX4032) drug treatment is prepared. Additionally, the potential of using CMP functionalized PEG hydrogels, which can provide better tunability is explored, to replace the existing invadopodia assay platform based on fluorescent gelatin.

Computational Optics of the Collagen Rich Tumor Microenvironment

Journal Article Computational Optics of the Collagen Rich Tumor Microenvironment Get access Kevin Eliceiri Kevin Eliceiri University of Wisconsin-Madison, Madison, Wisconsin, United States Search for other works by this author on: Oxford Academic Google Scholar Microscopy and Microanalysis, Volume 26, Issue S2, 1 August 2020, Page 1604, https://doi.org/10.1017/S1431927620018681 Published: 01 August 2020

3D collagen architecture regulates cell adhesion through degradability, thereby controlling metabolic and oxidative stress.

The collagen-rich tumor microenvironment plays a critical role in directing the migration behavior of cancer cells. 3D collagen architectures with small pores have been shown to confine cells and induce aggressive collective migration, irrespective of matrix stiffness and density. However, it remains unclear how cells sense collagen architecture and transduce this information to initiate collective migration. Here, we tune collagen architecture and analyze its effect on four core cell-ECM interactions: cytoskeletal polymerization, adhesion, contractility, and matrix degradation. From this comprehensive analysis, we deduce that matrix architecture initially modulates cancer cell adhesion strength, and that this results from architecture-induced changes to matrix degradability. That is, architectures with smaller pores are less degradable, and degradability is required for cancer cell adhesion to 3D fibrilar collagen. The biochemical consequences of this 3D low-attachment state are similar to those induced by suspension culture, including metabolic and oxidative stress. One distinction from suspension culture is the induction of collagen catabolism that occurs in 3D low-attachment conditions. Cells also upregulate Snail1 and Notch signaling in response to 3D low-attachment, which suggests a mechanism for the emergence of collective behaviors.

Nanoparticles for Targeting Intratumoral Hypoxia: Exploiting a Potential Weakness of Glioblastoma.

Extensive hypoxic regions are the daunting hallmark of glioblastoma, as they host aggressive stem-like cells, hinder drug delivery and shield cancer cells from the effects of radiotherapy. Nanotechnology could address most of these issues, as it employs nanoparticles (NPs) carrying drugs that selectively accumulate and achieve controlled drug release in tumor tissues. Methods overcoming the stiff interstitium and scarce vascularity within hypoxic zones include the incorporation of collagenases to degrade the collagen-rich tumor extracellular matrix, the use of multistage systems that progressively reduce NP size or of NP-loaded cells that display inherent hypoxia-targeting abilities. The unfavorable hypoxia-induced low pH could be converted into a therapeutical advantage by pH-responsive NPs or multilayer NPs, while overexpressed markers of hypoxic cells could be specifically targeted for an enhanced preferential drug delivery. Finally, promising new gene therapeutics could also be incorporated into nanovehicles, which could lead to silencing of hypoxia-specific genes that are overexpressed in cancer cells. In this review, we highlight NPs which have shown promising results in targeting cancer hypoxia and we discuss their applicability in glioblastoma, as well as possible limitations. Novel research directions in this field are also considered.

Heat-generating iron oxide nanocubes: subtle "destructurators" of the tumoral microenvironment.

Several studies propose nanoparticles for tumor treatment, yet little is known about the fate of nanoparticles and intimate interactions with the heterogeneous and ever-evolving tumor environment. The latter, rich in extracellular matrix, is responsible for poor penetration of therapeutics and represents a paramount issue in cancer therapy. Hence new strategies start aiming to modulate the neoplastic stroma. From this perspective, we assessed the efficacy of 19 nm PEG-coated iron oxide nanocubes with optimized magnetic properties to mediate mild tumor magnetic hyperthermia treatment. After injection of a low dose of nanocubes (700 μg of iron) into epidermoid carcinoma xenografts in mice, we monitored the effect of heating nanocubes on tumor environment. In comparison with the long-term fate after intravenous administration, we investigated spatiotemporal patterns of nanocube distribution, evaluated the evolution of cubes magnetic properties, and examined nanoparticle clearance and degradation processes. While inside tumors nanocubes retained their magnetic properties and heating capacity throughout the treatment due to a mainly interstitial extracellular location, the particles became inefficient heaters after cell internalization and transfer to spleen and liver. Our multiscale analysis reveals that collagen-rich tumor extracellular matrix confines the majority of nanocubes. However, nanocube-mediated hyperthermia has the potential to "destructure" this matrix and improve nanoparticle and drug penetration into neoplastic tissue. This study provides insight into dynamic interactions between nanoparticles and tumor components under physical stimulation and suggests that nanoparticle-mediated hyperthermia could be used to locally modify tumor stroma and thus improve drug penetration.

Relaxin treatment of solid tumors: effects on electric field–mediated gene delivery

Pulsed electric fields have been shown to enhance interstitial transport of plasmid DNA (pDNA) in solid tumors in vivo. However, the extent of enhancement is still limited partly due to the collagen component in extracellular matrix. To this end, effects of collagen remodeling on interstitial electrophoresis were investigated by pretreatment of tumor-bearing mice with a recombinant human relaxin (rh-Rlx). In the study, two tumor lines (4T1 and B16.F10) were examined and implanted s.c. to establish two murine models: dorsal skin-fold chamber (DSC) and hind leg. Effects of rh-Rlx on pDNA electrophoresis were measured either directly in the DSC model or indirectly in the hind leg model via reporter gene expression. It was observed that rh-Rlx treatment reduced collagen levels in the hind leg tumors but not in the DSC tumors. The observation correlated with the results from electromobility experiments, where rh-Rlx treatment enhanced transgene expression in 4T1 hind leg tumors but did not increase the electromobility of pDNA in the DSC tumors. In addition, it was observed that pDNA binding to collagen could block its diffusion in collagen gel in vitro. These observations showed that effects of rh-Rlx on the collagen content depended on microenvironment in solid tumors and that rh-Rlx treatment would enhance electric field-mediated gene delivery only if it could effectively reduce the collagen content in collagen-rich tumors.

Cancer Cell Death Enhances the Penetration and Efficacy of Oncolytic Herpes Simplex Virus in Tumors

The success of tumor oncolytic virotherapy is limited by the poor penetration of virus in tumors. Interstitial collagen fibers and the narrow spacing between cancer cells are major barriers hindering the movement of large viral particles. To bypass the cellular barrier, we tested the hypothesis that the void space produced by cancer cell apoptosis enhances the initial spread and efficacy of oncolytic herpes simplex virus (HSV). In mice with mammary tumors, apoptosis was induced by doxycycline-regulated expression/activation of CD8/caspase-8, paclitaxel, or paclitaxel plus tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). In both collagen-poor and collagen-rich tumors, apoptosis or necrosis increased the initial intratumoral spread of HSV. Compared with the isolated pattern of HSV infection generally located in the center of control tumors, apoptosis induction and a single i.t. injection of virus produced an interconnected and diffuse pattern of infection, which extended from the tumor center to the periphery. This interconnected pattern of viral infection correlated with the formation of void spaces and channel-like structures in apoptosis-rich tumor areas. We also show that the i.t. injection of HSV after caspase-8 activation or paclitaxel-TRAIL pretreatment retards tumor growth, whereas HSV administration before tumor cell death induction did not improve therapeutic efficacy. Hence, our findings show that the induction of cancer cell death before the injection of oncolytic HSV enhances intratumoral virus delivery/penetration and antitumor efficacy.