Periodic Reporting for period 2 - METALERT-STOP (Imaging, characterizing and targeting metastatic niches in melanoma)
Periodo di rendicontazione: 2022-06-01 al 2023-11-30
The main problem addressed in this proposal the mechanism(s) that underlie the high metastatic potential of cutaneous melanoma, the most aggressive form of skin cancer.
We chose melanoma precisely because its socioeconomical impact. First, this is a tumor of increasing incidence, with 1/35 individuals of fair skin expected to develop this malignancy during their lifetime. Secondly, melanoma is the only cancer where lesions barely above 1 mm in depth already bear high risk for metastasis. Large-scale analyses have identified a plethora of (epi)genetic alterations in aggressive melanomas. However, no genetic biomarker has yet been approved as a bona fide prognostic indicator. Thirdly, a key complication in this field has been the paucity of imaging tools to visualize premetastatic niches in vivo.
The overall objectives of this project are:
(1) When and how melanomas act “at a distance” preparing pre-metastatic niches before dissemination?
(2) How do melanoma cells evade the immune system?
(3) What is the impact of the microenvironment, particularly alarmins and bacteria-response genes?
(4) How to deactivate premetastatic niches pharmacologically?
We chose melanoma precisely because its socioeconomical impact. First, this is a tumor of increasing incidence, with 1/35 individuals of fair skin expected to develop this malignancy during their lifetime. Secondly, melanoma is the only cancer where lesions barely above 1 mm in depth already bear high risk for metastasis. Large-scale analyses have identified a plethora of (epi)genetic alterations in aggressive melanomas. However, no genetic biomarker has yet been approved as a bona fide prognostic indicator. Thirdly, a key complication in this field has been the paucity of imaging tools to visualize premetastatic niches in vivo.
The overall objectives of this project are:
(1) When and how melanomas act “at a distance” preparing pre-metastatic niches before dissemination?
(2) How do melanoma cells evade the immune system?
(3) What is the impact of the microenvironment, particularly alarmins and bacteria-response genes?
(4) How to deactivate premetastatic niches pharmacologically?
This project is aimed to define new mechanisms of melanoma progression and metastasis, with a particular focus on the interplay between tumor cells, the lymphatic vasculature and the immune system at early stages of the disease. We are also interested in the development of more efficient anticancer treatments.
Progress in this first period is as follows:
Aim 1. When and how melanomas act “at a distance” preparing pre-metastatic niches before dissemination?
We have previously identified the growth factor Midkine (MDK) as a melanoma-secreted protein with potent pro-metastatic and immune suppressive functions. Using single cell RNA seq and validating results by immunohistochemistry and flow cytometry, we have now identified a variety of cell types (particularly CD8 T cells) modulated in a systemic manner by MDK at early time points of melanoma development. Gene sets were identified that defined overall patient survival and response to immune checkpoint blockade (Contreras et al. in preparation).
Aim 2. How do melanoma cells evade the immune system and whether these mechanisms are equivalent at different anatomical sites?
Combining cellular systems, mouse models, and clinical datasets, we have now identified MDK as a potent repressor of dentritic cell (DC) differentiation, activation, and function. Moreover, a MDK-associated signature in DCs was found to define bad prognosis and resistance to ICB in human patients (Catena et al. BioRxiv 2022; Catena et al. in revision).
Aim 3. What is the impact of the microenvironment, particularly alarmins and bacteria-response genes?
Performing RNA sequencing in MDK expressing and lacking tumor implants in mice have identified a putative new role of this protein in the control of various alarmins. This function was found linked to associated with a marked rewiring of the immune system, particularly CD8 T cells and macrophages.
Aim 4. How to deactivate premetastatic niches (with dsRNA mimics and strategies to blunt selective vulnerabilities)?
We have published an article on how to deactivate MDK therapeutically with dsRNA nanoplexes (Olmeda et al. EMBO Mol Med 2021). Here is a summary that highligts main basic clinical implications of this study:
“Here we exploit immunocompetent and immunodeficient mouse models for live imaging of Vegfr3-driven neolymphangiogenesis, as a versatile platform for drug screening in vivo. Spatio-temporal analyses of autochthonous melanomas and patient-derived xenografts identified double-stranded RNA mimics (dsRNA nanoplexes) as potent inhibitors of neolymphangiogenesis, metastasis and post-surgical disease relapse. Mechanistically, dsRNA nanoplexes were found to exert a rapid dual action in tumor cells and in their associated lymphatic vasculature, involving the transcriptional repression of the lymphatic drivers Midkine and Vegfr3, respectively. This suppressive function was mediated by a cell-autonomous type I interferon signaling and was not shared by FDA-approved anti-melanoma treatments. These results reveal an alternative strategy for targeting the tumor cell-lymphatic crosstalk and underscore the power of Vegfr3-lymphoreporters for pharmacological testing in otherwise aggressive cancers.”
Progress in this first period is as follows:
Aim 1. When and how melanomas act “at a distance” preparing pre-metastatic niches before dissemination?
We have previously identified the growth factor Midkine (MDK) as a melanoma-secreted protein with potent pro-metastatic and immune suppressive functions. Using single cell RNA seq and validating results by immunohistochemistry and flow cytometry, we have now identified a variety of cell types (particularly CD8 T cells) modulated in a systemic manner by MDK at early time points of melanoma development. Gene sets were identified that defined overall patient survival and response to immune checkpoint blockade (Contreras et al. in preparation).
Aim 2. How do melanoma cells evade the immune system and whether these mechanisms are equivalent at different anatomical sites?
Combining cellular systems, mouse models, and clinical datasets, we have now identified MDK as a potent repressor of dentritic cell (DC) differentiation, activation, and function. Moreover, a MDK-associated signature in DCs was found to define bad prognosis and resistance to ICB in human patients (Catena et al. BioRxiv 2022; Catena et al. in revision).
Aim 3. What is the impact of the microenvironment, particularly alarmins and bacteria-response genes?
Performing RNA sequencing in MDK expressing and lacking tumor implants in mice have identified a putative new role of this protein in the control of various alarmins. This function was found linked to associated with a marked rewiring of the immune system, particularly CD8 T cells and macrophages.
Aim 4. How to deactivate premetastatic niches (with dsRNA mimics and strategies to blunt selective vulnerabilities)?
We have published an article on how to deactivate MDK therapeutically with dsRNA nanoplexes (Olmeda et al. EMBO Mol Med 2021). Here is a summary that highligts main basic clinical implications of this study:
“Here we exploit immunocompetent and immunodeficient mouse models for live imaging of Vegfr3-driven neolymphangiogenesis, as a versatile platform for drug screening in vivo. Spatio-temporal analyses of autochthonous melanomas and patient-derived xenografts identified double-stranded RNA mimics (dsRNA nanoplexes) as potent inhibitors of neolymphangiogenesis, metastasis and post-surgical disease relapse. Mechanistically, dsRNA nanoplexes were found to exert a rapid dual action in tumor cells and in their associated lymphatic vasculature, involving the transcriptional repression of the lymphatic drivers Midkine and Vegfr3, respectively. This suppressive function was mediated by a cell-autonomous type I interferon signaling and was not shared by FDA-approved anti-melanoma treatments. These results reveal an alternative strategy for targeting the tumor cell-lymphatic crosstalk and underscore the power of Vegfr3-lymphoreporters for pharmacological testing in otherwise aggressive cancers.”
AIM 1. PREMETASTATIC NICHE FORMATION
We have performed a comprehensive analysis of premetastatic niches in melanoma. These studies identified unexpected immune suppressive roles of MDK at early stages of melanoma progression.
AIM 2. NEW MECHANISMS OF IMMUNEMODULATION IN MELANOMA
We have discovered a new role of MDK as a potent immune suppressor of dendritic cell differentiation and function, that indeed will open new avenues of research not only for melanoma, but for a broad spectrum of cancer types where this protein is overexpressed.
Papers are as follows:
1. Catena X, Contreras-Alcaide M, Cerezo-Wallis D, Larrea Naiara J, Olmeda D, Calvo GT, Mucientes C, Oterino S and Soengas MS. Systemic effects of melanoma-secreted MIDKINE in the inhibition of dendritic cell differentiation and function. BioRxiv. doi: https://doi.org/10.1101/2022.12.28.521901(si apre in una nuova finestra)
2. Catena X, Contreras-Alcaide M, Cerezo-Wallis D, Larrea Naiara J, Olmeda D, Calvo GT, Suárez J., Mucientes C, S-Frago., Oterino S. Martínez L, Megías D, Sancho D, Seretis A., Dudziak D, Stoitzner P. and Soengas MS. Melanoma-secreted Midkine rewires dendritic cells systemically impairing immune surveillance and compromising immune checkpoint blockade (Submitted)
AIM 3. MICROENVIRONMENTAL FACTORS IN THE CONTROL OF MELANOMA PROGRESSION BY MDK
We have generated a new mouse reporter mouse model for MDK (Mdk-IRES-Luc2-tdTtomato) which allows us to visualize Mdk mRNA expression in vivo, (i) during embryonic development,(ii) prior/post tumor implantation and (iii) before and after treatment with anticancer agents. These animals are being tested to define feedback loops between melanoma cells and the stromal and vascular microenvironment, with a particular emphasis on various alarmins and lipid-metabolism controllers (Olmeda et al. in preparation). We expect these animals to serve as a platform for the discovery of yet novel pro-metastatic and immunomodulatory factors.
AIM 4. METALERT MICE FOR DRUG SCREENING AND IDENTIFICATION OF POTENT ANTI-METASTATIC AGENTS.
Our studies have identified dsRNA mimics (the polyplex BO-110) as potent dual inhibitors of MDK and the lymphatic factor VEGFR3. These functions are distinct from the reported action of lymphangiogenic factors in clinical testing and were not recapitulated by FDA-approved therapies based on BRAF inhibition and checkpoint blockade. An additional unexpected result of this study was the finding that one single administration of BO-110 nearly abrogated Vegfr3Luc emission in vivo in melanomas of different genetic backgrounds. This information could potentially be used in clinical trials that are currently testing derivatives of BO-110 and other formulations of dsRNA.
Paper is as follows:
3. Olmeda D, Cerezo-Wallis D, Mucientes C, Calvo TG, Cañón E, Alonso-Curbelo D, Ibartz N, Muñoz J, Rodríguez-Peralto JL, Ortiz-Romero P, Ortega S and Soengas MS. Live imaging of neolymphangiogenesis identifies acute antimetastatic roles of dsRNA mimics (2021). EMBO Mol Med. Dec 7;13(12):e12924. doi: 10.15252/emmm.202012924.
We have performed a comprehensive analysis of premetastatic niches in melanoma. These studies identified unexpected immune suppressive roles of MDK at early stages of melanoma progression.
AIM 2. NEW MECHANISMS OF IMMUNEMODULATION IN MELANOMA
We have discovered a new role of MDK as a potent immune suppressor of dendritic cell differentiation and function, that indeed will open new avenues of research not only for melanoma, but for a broad spectrum of cancer types where this protein is overexpressed.
Papers are as follows:
1. Catena X, Contreras-Alcaide M, Cerezo-Wallis D, Larrea Naiara J, Olmeda D, Calvo GT, Mucientes C, Oterino S and Soengas MS. Systemic effects of melanoma-secreted MIDKINE in the inhibition of dendritic cell differentiation and function. BioRxiv. doi: https://doi.org/10.1101/2022.12.28.521901(si apre in una nuova finestra)
2. Catena X, Contreras-Alcaide M, Cerezo-Wallis D, Larrea Naiara J, Olmeda D, Calvo GT, Suárez J., Mucientes C, S-Frago., Oterino S. Martínez L, Megías D, Sancho D, Seretis A., Dudziak D, Stoitzner P. and Soengas MS. Melanoma-secreted Midkine rewires dendritic cells systemically impairing immune surveillance and compromising immune checkpoint blockade (Submitted)
AIM 3. MICROENVIRONMENTAL FACTORS IN THE CONTROL OF MELANOMA PROGRESSION BY MDK
We have generated a new mouse reporter mouse model for MDK (Mdk-IRES-Luc2-tdTtomato) which allows us to visualize Mdk mRNA expression in vivo, (i) during embryonic development,(ii) prior/post tumor implantation and (iii) before and after treatment with anticancer agents. These animals are being tested to define feedback loops between melanoma cells and the stromal and vascular microenvironment, with a particular emphasis on various alarmins and lipid-metabolism controllers (Olmeda et al. in preparation). We expect these animals to serve as a platform for the discovery of yet novel pro-metastatic and immunomodulatory factors.
AIM 4. METALERT MICE FOR DRUG SCREENING AND IDENTIFICATION OF POTENT ANTI-METASTATIC AGENTS.
Our studies have identified dsRNA mimics (the polyplex BO-110) as potent dual inhibitors of MDK and the lymphatic factor VEGFR3. These functions are distinct from the reported action of lymphangiogenic factors in clinical testing and were not recapitulated by FDA-approved therapies based on BRAF inhibition and checkpoint blockade. An additional unexpected result of this study was the finding that one single administration of BO-110 nearly abrogated Vegfr3Luc emission in vivo in melanomas of different genetic backgrounds. This information could potentially be used in clinical trials that are currently testing derivatives of BO-110 and other formulations of dsRNA.
Paper is as follows:
3. Olmeda D, Cerezo-Wallis D, Mucientes C, Calvo TG, Cañón E, Alonso-Curbelo D, Ibartz N, Muñoz J, Rodríguez-Peralto JL, Ortiz-Romero P, Ortega S and Soengas MS. Live imaging of neolymphangiogenesis identifies acute antimetastatic roles of dsRNA mimics (2021). EMBO Mol Med. Dec 7;13(12):e12924. doi: 10.15252/emmm.202012924.