Melanoma is the most lethal skin cancer, leading to an estimated 8,290 deaths and 100,640 new cases in 2024. Melanoma was traditionally very difficult to treat, which has changed in the past 15 years with the onset of immunotherapies. Despite their success, many patients do not respond to immunotherapy and are in need of secondary treatments. Different forms of cell death are able to improve the recognition of tumors by the immune system. A form of cell death called pyroptosis is when proteins in the cells, gasdermins, poke holes on the cell surface, causing various cellular components to leak out and stimulate the immune system. Pyroptosis improves immunotherapy in several cancers, thus we wondered if a particular form of pyroptosis driven by gasdermin D might improve the success of immunotherapy in melanoma. 

In the first aim of our project, we propose to study if causing pyroptosis through gasdermin D is able to effect tumor growth in cutaneous melanoma like model system. We will test if the immune system is needed for any tumor growth effects we observe after causing pyroptosis. Next, we will dissect the specific immune responses caused by gasdermin D driven pyroptosis, specifically testing the reliance of these immune responses on a specific population of immune stimulatory cells. Lastly, we will determine if specific factors released from pyroptotic cancer cells are driving the immune responses and tumor growth effects we observe. We ultimately expect to find that pyroptosis through gasdermin D slows tumor growth by engaging the immune system. 

For the second aim of this grant, we propose testing if pyroptosis induction through gasdermin D can improve the effect of immunotherapy in cutaneous melanoma tumor models. Specifically, we will investigate the best timing of pyroptosis induction to improve immunotherapy and lead to immune responses that can impact distant tumors that model metastases. We will also work with The Wistar Institute to screen thousands of anti-cancer drugs to discover which can cause pyroptosis through gasdermin D. We will then test to see if the drugs identified can slow the growth of primary and metastatic tumors through so called abscopal effects. With this aim we expect to leverage pyroptosis to improve immunotherapy in melanoma and discovery new treatments for melanoma. 

Immunotherapy has greatly improved survival for people with melanoma, but many patients also experience harmful side effects when the immune system attacks healthy tissues. Skin inflammation is among the most common problems, sometimes lowering quality of life or even discontinuing treatment.  

A growing body of research shows that neutrophils, a type of immune cell, play a central role in causing some of these toxicities. In our earlier studies in mice, we found that skin inflammation during immunotherapy was linked to neutrophils that express a molecule called VISTA. These findings suggest that VISTA-positive neutrophils may be especially important drivers of skin damage.  

We are now turning to human studies to determine whether the same process occurs in people. By examining skin samples from melanoma patients who developed immune-related side effects, we will test the role of VISTA-positive neutrophils in driving inflammation. Insights from this work may guide the development of safer and more potent immunotherapies that maintain strong anti-cancer activity while reducing harmful side effects.  

Immunotherapies including immune checkpoint blockade (ICB) have seen remarkable success in the clinic. Unfortunately, many patients with late-stage melanoma still exhibit only a partial response to therapy, where tumor lesions shrink temporarily but then continue to progress. If we could convert these partial responders to complete responders, we could make significant impacts on patient survival rates long-term. What causes these patients to differ from complete responders? If we think of the race between the growth and death of tumor cells, immunotherapy’s goal is to increase the death rate to overtake growth. In these partial responder patients, clearly the growth rate manages to catch up to and surpass the death rate and the race is eventually lost. We hypothesize that tumors can be divided into different types of regions that exhibit different levels of tumor cell death following ICB treatment. Some regions are “reactive” (RE) which exhibit enhanced levels of tumor cell death and others are “non-reactive” (NRE) which exhibit decreased levels of tumor cell death. The balance and interplay between these types of regions ultimately determines the outcome of therapy. How are these regions arranged in melanomas during ICB treatment? What causes these regions to develop? And if we look for these regions in patient samples, can we more accurately predict their treatment outcome. Our proposal will directly address these questions using advanced microscopy and spatial technologies. We will use mouse models of melanoma engineered to light up fluorescently when undergoing processes such as death, growth and quiescence. By imaging large pieces of these tumors, we will be able to precisely count the number of cells undergoing those processes and map their distribution in the spatial coordinates of the tumor. Once we have maps of the spatial distributions of tumor cell death, we will apply novel technologies to zoom in on the RE and NRE regions and ask, what types of cells are located there and how are they interacting? What kind of cell neighborhood leads to better or worse tumor cell elimination? 

We will then look at samples taken from patient tumors who underwent ICB therapy to examine if similar regions and features from our previous aims are found in these tissues. This will allow us to predict which region type (RE vs NRE) is more dominant as treatment progresses and thus the ultimate outcome of treatment on a per-patient basis. In other words, we can monitor the race between tumor death and growth in patients undergoing ICB and check our predictions against the known clinical outcome. We envision our work will reveal additional vulnerabilities to target in melanoma in combination with ICB to increase survival in patients. Additionally, the prognostic model we generate will provide an invaluable tool in the clinic for monitoring and predicting the outcome of ICB in patients, allowing for more precise and personalized cancer care. 

Skin cancer is one of the most common types of cancer, with melanoma being the deadliest form. In the United States, approximately 104,960 new cases of melanoma are expected in 2025, and about 8,500 people are projected to die from the disease. Melanomas often result from genetic mutations in genes controlling cell growth. A key pathway involved in this is the MAPK pathway (Mitogen-Activated Protein Kinase pathway), which transmits growth signals from the cell surface to the nucleus. When mutated, this pathway becomes overactive, leading to uncontrolled cell growth and melanoma. The BRAF (V600E) mutation is one of the most common causes, leading to constant activation of MAPK and uncontrolled cell division. Treating advanced melanoma, especially when it spreads to other parts of the body (metastatic melanoma), was historically difficult. Over the past decade, targeted therapies have changed this. BRAF inhibitors were developed to block the BRAF (V600E) mutation and paradoxically hyperactivate the MAPK pathway in normal cells, preserving healthy tissue function. This effect makes the combination of BRAF inhibitors and MEK inhibitors (which block another part of the MAPK pathway) more tolerable, improving patient survival and quality of life. However, some patients eventually develop resistance, causing cancer progression. Ongoing research (including our) focuses on finding ways to overcome it. Also, around 20% of melanoma cases carry mutations in the NRAS gene, which also overactivates the MAPK pathway. However, unlike BRAF-mutant melanoma, there are no approved targeted therapies for NRAS-mutant melanoma, and BRAF inhibitors are ineffective. To address this, a new class- pan-RAF inhibitors were developed to block RAF proteins in all contexts. However, these inhibitors shut down the MAPK pathway in both cancer and healthy cells, causing significant side effects. Combining pan-RAF inhibitors with MEK inhibitors improves their cancer-fighting ability but still causes unwanted suppression of normal cell function. As a result, treatment options for NRAS-mutant melanoma remain limited and urgently needed. 

Here, we propose a novel approach to melanoma treatment by developing and characterizing a new class of RAF inhibitors targeting all classes of melanomas. Like BRAF inhibitor, but with a distinct mechanism, this class of RAF inhibitor selectively increase MAPK pathway activity in healthy cells while blocking the pathway in all melanoma cells we tested. This approach allows us to combine these RAF inhibitors with MEK inhibitors (and other MAPK inhibitors) to enhance cancer cell potency without affecting normal cells. Using this cutting-edge approach, we will provide new insight into the molecular mechanism of action underlying this novel class of RAF inhibitor. This project has the potential to advance novel drug combinations into human clinical trials, offering new hope for patients with BRAF-mutant, NRAS-mutant, and treatment-resistant melanoma. 

Acral and mucosal melanomas(A&MM) are rare but aggressive types of melanoma, making up less than 5% of all cases. Unlike the more common type of melanoma, which develops on sun-exposed skin, A&MM arise in areas not typically exposed to the sun—such as the palms, soles, under the nails (acral melanoma), or in mucous membranes like the mouth, nose, and genitals (mucosal melanoma). These cancers follow a different genetic path and do not respond well to standard treatments, making them particularly difficult to treat. Research has shown that many A&MM tumors have mutations that activate a key cellular pathway known as the RAS signaling pathway, which drives cancer growth. However, there are currently no effective targeted treatments for these patients.

A new class of drugs, called RAS(ON) inhibitors, has been developed to block the activity of RAS, offering a potential breakthrough for patients with RAS-driven cancers. Early studies suggest that these drugs could be effective in slowing tumor growth. In my lab research, I tested a RAS(ON) inhibitor on tumor models derived from two patients—one with acral melanoma and one with mucosal melanoma. While the tumors initially shrank in response to the drug, which is a great win for this challenging cancer type, they later became resistant, meaning the treatment stopped working. This indicates that while RAS(ON) inhibitors hold great promise, we need to understand how resistance develops and find ways to prevent or overcome it. This study has two main goals. First, I will test RAS(ON) inhibitors on a larger set of patient-derived tumor models grown in mice to determine which types of A&MM are most likely to respond. Second, I will investigate how resistance to these drugs develops by analyzing genetic changes in tumors before and after they stop responding. By identifying the biological mechanisms that allow the cancer to escape treatment, I aim to discover combination therapies that can enhance the effectiveness of RAS(ON) inhibitors. The results of this research will provide valuable insights into how A&MM can be better treated, guiding the development of new, more effective therapies. Ultimately, this work aims to bring new hope to patients with these rare and challenging melanomas by advancing precision medicine approaches tailored to their unique cancer biology. 

Prolonged sun exposure combined with other multi-factorial cues can transform skin’s melanocytes into melanoma cells characterized by uncontrolled proliferation. If not removed on time, melanoma cells can quickly spread into other tissues and form lethal secondary tumors called metastasis. Several anti-melanoma drugs have been developed in the last decade, in particular immunotherapy to help our natural protective barrier or immune system to eradicate cancer cells. Immune system is a group of diverse cells able to activate biological responses to eliminate foreign organisms (i.e., viruses) or abnormal cells including cancer cells. However, melanoma cells can modify the expression of genes that confer them special “identities” allowing them to become invisible to the immune system. Therefore, a better characterization of these identities remains essential to define new therapeutic targets. Using complex bioinformatic analyses, we compared the expression of several genes among different tumors and normal tissues. Surprisingly, we identified a new gene named HOXD13 almost exclusively expressed in melanoma cells of patients non responsive to immunotherapy. Through genetically modified mice able to develop melanoma, we observed that HOXD13 activation confers to melanoma cells an invisible identity to the immune system. Using other molecular techniques, we demonstrate that HOXD13 activates two other genes named CD73 and NGFR which we suspect being the main actors of immune invisibility. Altogether, our studies will reveal whether HOXD13/CD73/NGFR axes inhibition could make melanoma cells more visible to the immune system and improving immunotherapy response with a direct relevance for the patients.

When cancer cells metastasize to the brain, the diagnosis becomes terminal. In melanoma brain metastasis, patients are only given months to live. There is an urgent need to develop treatment strategies to improve clinical outcomes for these patients. Although immunotherapy has been successfully used to treat some types of cancers, its effectiveness is limited in many solid tumors, including brain metastasis. One reason for this is due to the accumulation of myeloid cells in the tumor. These myeloid cells come from bone marrow, and enter the circulation when tumors produce recruitment signals into the blood. One of the main functions of these myeloid cells is to suppress the killing activity of T cells, allowing the tumors to grow while evading the body’s natural immune system. In the brain, there are specialized myeloid cells called microglia. In brain tumors, these microglia are thought to have both unique and shared functions to the circulating myeloid cells. Therefore, a better understanding of these microglia and how they may contribute to brain tumor progression will lead to the development of new therapies. This study will investigate a biological signaling pathway that is found to be activated in microglia as a response to invading cancer cells. This pathway is part of the natural immune response exerted by microglia in many other nervous system diseases, such as Alzheimer’s and multiple sclerosis. In these other diseases, the activated microglia play a role in resolving inflammation, which has beneficial effects by reducing damage to the local organ. In cancer, however, this has the opposite effect and protects the tumor, allowing it to grow unchecked. In this study, we propose to investigate this pathway and its involvement in brain tumor progression. Completion of this project will establish targeting of microglia as a new therapeutic approach, in combination with immunotherapies, for brain metastasis patients.

Uveal Melanoma (UM), a deadly cancer arising in the eye, is the most common eye cancer in adults. Early detection of UM is important due to the cancer’s ability to spread to the rest of the body early and because effective treatments are available to reduce its spread. Despite the availability of effective treatments, more than half of patients’ cancer spreads to the rest of the body, suggesting that UM may spread before the time of treatment. Choroidal nevi are benign tumors that are commonly seen in patients’ eyes and rarely can turn into cancer. Choroidal nevi can look like UM, which makes the diagnosis of these eye tumors challenging. Therefore, there is a need to identify and treat small UM to minimize the number of tumors that are observed and subsequently grow during the observation period. However, current screening methods face inherent limitations, particularly in regions with limited access to specialized eye cancer doctors. Machine learning (ML) is a field of study in artificial intelligence that can be used to assist in disease diagnosis. ML offers a promising approach to improve the identification and evaluation of eye tumors, thereby providing a potential tool for eye doctors both in the community and who specialize in eye cancer. Despite the significant research being done with ML in medical imaging, few studies have worked towards an ML tool for eye tumors like choroidal nevi and UM. Our previous work used ML to screen images of patients’ eyes to successfully find choroidal nevi and UM. We also used ML to evaluate images and ultrasound of patients’ eyes to assess choroidal nevi and UM for their ability to spread to the rest of the body. The objective of this proposed project is to assess the ability of ML to diagnose choroidal nevi and UM. We will use a large collection of images from the University of Illinois Chicago, which includes multiple different types of images taken of many patients with choroidal nevi and UM. We propose to develop and then improve ML tools for the screening and diagnosis of choroidal nevi and UM. We will use additional information about the patient’s medical history to help the tools perform better. We will also annotate the images before providing them to the ML tools to see if this improves the performance in screening and diagnosing choroidal nevi and UM. After identifying the best-performing ML tool in this research study, we will test this tool to perform other tasks related to the choroidal nevi and UM. We will test the ML tool’s performance in predicting the likelihood that a choroidal nevus will grow and the likelihood that a UM will spread to the rest of the body. We expect these results to provide a better understanding of how ML can be developed into a clinically useful tool to inform and guide management decisions for choroidal nevi and UM in community eye clinics and eye cancer specialist clinics to potentially save patient lives.

A major reason that T cell-based melanoma immunotherapies can fail, either initially or after a period of treatment, is due to mutations that result in CD8+ T cells being incapable of directly recognizing and killing tumor cells. Many melanomas, however, express HLA-II which enables tumor cells to be recognized and eliminated by an alternative lymphocyte subset called CD4+ T cells. Unlike tumor cell killing by CD8+ T cells, the molecular mechanisms governing response and resistance of tumor cells to CD4+ T cell killing have previously not been studied in a systematic and unbiased fashion. This gap in knowledge limits the rational application of CD4+ T cells as an effector population in melanoma immunotherapies. Using a genome-scale CRISPR screen I recently identified genes that confer enhanced sensitivity or resistance of melanoma cells to CD4+ cell killing. I discovered that loss of function of the RNA-binding protein PCBP2 led to enhanced killing of tumor cells by CD4+ T cells. The tumor-intrinsic role of PCBP2 in tumor immunology has previously not been studied. In my project, I seek to define the molecular mechanisms that control enhanced tumor-intrinsic sensitivity of PCBP2-deleted melanoma cells to immunologic attack by CD4+ T cells. These findings will inform a novel, mechanism-based immunotherapeutic approach that augments antitumor efficacy of CD4+ T cells in melanoma and other common cancers.

Metastatic melanoma is one of the most aggressive, and complex cancers with a 35% 5-year survival rate. Therapies using T cells that are genetically engineered to express tumor-recognizing molecules, known as chimeric antigen receptors (CAR), have shown promise against some blood cancers, but not in solid tumors like melanoma. The proposed study aims to engineer CAR T cells to overcome the suppressive tumor microenvironment and engage other immune cells to mount a more effective therapeutic response. This will be accomplished by creating combination immunotherapies through developing CAR T cells that produce drugs that engage other immune cells and help develop immunological memory to the tumor. The overall goal of our research is to develop optimized immune cell therapies for patients with melanoma by understanding how these treatments interact with the tumor environment and patients’ immune system.