Immunotherapy

RET or rearranged during transfection

Tyrosine kinase inhibitors (TKIs)

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Immunogenic peptide priming of dendritic cells for RET+ NSCLC

2024

Partner Awards

Grant title (if any)

The Hamoui Foundation/LUNGevity Lung Cancer Research Award Program

Amy Cummings, MD, PhD

University of California, Los Angeles

Los Angeles

This project will explore the use of neoantigens to evaluate immunogenic priming of dendritic cells (DC) in RET+ NSCLC. Neoantigens are short protein fragments present only in cancer cells that bind to genetically encoded proteins known as human leukocyte antigens (HLA). Dr. Cummings will use features of HLA to predict which cancer-specific protein fragments best match an individual’s immune system, utilizing a biobank of RET-rearranged NSCLC biospecimens. This approach could help identify optimal immunogenic targets, that could be translated into a pathway for clinical use of personalized DC vaccines.

Research Summary

RET-rearranged non-small cell lung cancer (NSCLC) is a rare subtype of lung cancer that is driven by growth signals triggered by RET activation. RET-specific inhibitors are effective initially, but most benefit from this treatment for only 1-2 years before additional treatment is needed. Chemotherapy is a widely-available option but typically provides less than six months of benefit, and it is unclear whether immunotherapy alone or in combination with chemotherapy is a better option. Findings from other gene-rearranged NSCLC studies, particularly those on ALK-rearranged NSCLC, suggest that immunotherapy works better when the immune system is better exposed to abnormalities created by the gene-rearrangement. These are neoantigens, or short protein fragments present only in cancer cells that bind to human leukocyte antigen (HLA), a scaffold that displays these protein fragments to the immune system. One issue with this approach is that these fragments have to be specifically matched to the immune system of an individual, and even the most common forms of HLA are only found in 20% of people. This means that these types of approaches would be applicable to at most 1 out of 5 people with RET-rearranged NSCLC. Our techniques broaden this approach by using features of HLA to predict which cancer-specific protein fragments best match an individual’s immune system (motif neoepitopes), including neoantigens from RET rearrangements and those predicted from the individual’s tumor. We propose to use our biobank of RET-rearranged NSCLC biospecimens, which have not been previously analyzed, to determine whether we can detect and elicit enhanced immune responses with motif neoepitopes, neoantigens related to RET-rearrangements, or other predicted neoantigens. We can then offer this approach in a currently open clinical trial investigating immune system optimization through an application to the FDA.

Technical Abstract

RET-rearranged non-small cell lung cancer (NSCLC) presents challenges in management following progression on selective tyrosine kinase inhibitors (TKIs). Platinum-based chemotherapy and docetaxel are available options but are without durable benefit. Real world data with single-agent and combination chemo-immunotherapy suggests modest benefit and possible efficacy if immunotherapy-based approaches are appropriately optimized. For the past decade, our group has meticulously curated hundreds of NSCLC biospecimens including matched tissue and blood from multiple timepoints, including 7 RET-rearranged NSCLC cases that have not previously been analyzed. We have extensive expertise in neoepitope prediction and personalized immunotherapy through dendritic cell (DC)-based vaccination. Our most recent collaboration enabled functional assessments of T-cells through nanovial-based affinity repertoires, further enhancing our ability to predict and translate immunogenic peptides through a personalized vaccine-based program. We propose to use our RET-rearranged NSCLC biospecimens to systematically study T-cell-specific responses to identify optimal immunogenic peptide targets, an approach that could be translated in our currently open and approved DC vaccination trial (NCT03546361) through single patient exemptions.

Key words

RET or rearranged during transfection

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Targeting tumor associated macrophages in immunotherapy resistant NSCLC

2024

Partner Awards

Grant title (if any)

Brown/LUNGevity Award to Understand Mechanisms of Resistance to Immunotherapy

Dwight Owen, MD, MSc

The Ohio State University

Columbus

This project will investigate the role of cells called macrophages, key components of the immune system that have multiple functions, including immune surveillance within a unique communication pathway called hedgehog (Hh). The hedgehog signaling pathway is involved in cell growth and differentiation, as well as maintenance of stem cells and tissue repair. Disruption or inhibition of Hh can create an environment that is less favorable for survival of cancer cells, allowing a patient’s immune system to combat it more effectively. This research has the potential to benefit patients who have been diagnosed with NSCLC, who have not responded to current treatments including immunotherapy by boosting the body’s own defense mechanisms.

Research Summary

Lung cancer remains one of the most lethal types of cancer worldwide, with non-small cell lung cancer (NSCLC) accounting for a majority of cases. The goal of our research is to better understand the relationship between certain immune cells called macrophages and NSCLC, and how this interaction contributes to the cancer's survival and resistance to treatment. The scientific premise of our project lies in investigating a unique communication pathway known as hedgehog signaling within these macrophages and determining how it impacts the immune system's ability to fight lung cancer. If successful, our research has the potential to benefit patients who have been diagnosed with NSCLC, particularly those who have not responded to current treatments including treatment with immune therapies. By disrupting the hedgehog signaling pathway in macrophages, we hope to create a tumor immune environment that is less favorable for cancer cell survival, allowing patients' immune systems to effectively combat the disease. This research can pave the way for innovative therapeutic approaches that boost the body's own defense mechanisms.

Technical Abstract

The prognosis for patients with metastatic non-small cell lung cancer (NSCLC) remains poor despite recent progress in immune checkpoint blockade (ICB) therapy. Thus, there is an urgent need to understand mechanisms for lung cancer immune evasion within the tumor microenvironment (TME) in order to develop more effective and durable strategies for treating lung cancer. Tumor associated macrophages (TAMs), a major component of the tumor stromal mass, generally display an anti-inflammatory phenotype and can facilitate tumor growth by promoting angiogenesis, invasion, and metastasis, as well as immune evasion. However, it remains largely undefined exactly how these TAMs regulate anti-tumor immune responses within the TME. Will test the hypothesis that hedgehog signaling in TAMs interferes with recruitment of CD8+ T cells to the TME through the following specific aims: 1) Investigate the role of Hh inhibition with anti-PD-L1 therapy in non-small cell lung cancer; 2) Study the impact of Hh inhibition on TAMs and changes within the TME. The objective of this project is to understand signals required for functional polarization of TAMs within the TME and its contributions to immune cell dysregulation and cancer progression, and whether combined Hh inhibition and ICB can overcome resistance to immunotherapy.

Key words

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Developing EGFRxHER3 bispecific CAR-T cells for targeting EGFR TKI DTPCs

2024

Career Development Award

Yan Yang, PhD

MD Anderson Cancer Center

Houston

In patients with EGFR-mutant NSCLC, tyrosine kinase inhibitors (TKIs) have been an effective treatment, but over time these patients develop resistance to TKIs, leading to tumor relapse. Dr. Yang’s project focuses on cancer cells called drug-tolerant persisters (DTPs), which are implicated in TKI resistance. A gene called HER3 is expressed in DTPs, and Dr. Yang will use specially engineered immune cells, called CAR-T cells, to target both HER3 and EGFR simultaneously. If successful, this approach would result in a bi-specific CAR-T cell that can be further evaluated in clinical trials.

Research Summary

In lung cancer patients with a specific genetic mutation in EGFR, certain drugs can be helpful, but over time, the cancer can learn to resist these drugs, making them less effective. We're focusing on a small group of tough cancer cells, called DTPCs, that survive the initial treatment because they might be the key to curing the cancer for good. We've noticed that a gene called HER3 is expressed more in these cells. We also have promising results from CAR-T cell therapy. It is a treatment using T cells, a type of immune cells, engineered with the chimeric antigen receptor (CAR) so they can find and destroy cancer cells. We've found that CAR-T cells targeting EGFR can kill DTPCs. We believe if we use these T cells to attack both HER3 and EGFR at the same time, we might have a better chance of killing off these stubborn cancer cells. To make sure this idea works, we'll first check if HER3 is indeed more expressed in these DTPCs in the lab and samples from patients. Then, we'll test our HER3-targeting CAR-T cells to see if they can kill these DTPCs. If that looks promising, we'll tweak the T cells to target both HER3 and EGFR and see if they work even better. If all goes well, we'll be able to make one of the first effective CAR-T cells to target both HER3 and EGFR and try them out in clinical trials.

Technical Abstract

In NSCLC patients harboring mutant EGFR, treatment with tyrosine kinase inhibitors (TKIs) has demonstrated efficacy. However, the emergence of resistance to EGFR TKIs remains a significant challenge, leading to disease progression. Targeting drug-tolerant persister cells (DTPCs), a rare subpopulation surviving initial treatment, emerges as a more effective strategy than awaiting the development of complete drug resistance, holding potential for curative cancer therapy. Based on our preliminary data on the upregulation of HER3 in DTPCs resistant to EGFR TKIs and the potent antitumor activity of EGFR-targeting chimeric antigen receptor (CAR)-T cells against DTPCs. We propose that HER3 is a promising target expressed on EGFR-TKI DTPCs and that CAR-T cells simultaneously targeting both EGFR and HER3 are an effective approach for targeting DTPCs with improved overall efficacy. In this proposal, we will evaluate HER3 as a therapeutic target for CAR-T cell therapy against EGFR-TKI DTPCs by validating its expression in DTPCs using in vitro cell models, in vivo xenograft and PDX models, and patient tissues. Additionally, we will evaluate the anti-tumor activity of HER3-targeting CAR-T cells against DTPCs. Subsequently, we will develop and characterize EGFRxHER3 bispecific CAR-T cells, evaluating their CAR expression, antigen binding affinity, and anti-tumor activity. We next will assess their anti-tumor efficacy in xenograft and PDX models to guide the selection of the most promising bispecific CAR-T cells for further development. If completed successfully, we will have EGFRxHER3 bispecific CAR-T cells ready for GMP-compliant clinical-grade CAR manufacturing, in preparation for clinical trial evaluation.

Key words

Epidermal growth factor receptor (EGFR)

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Building Reliable Oncology Navigation to Ensure Adjuvant Management: BRONx-TEAM Project

2024

Career Development Award

Tamar Nobel, MD, MPH

Montefiore Medical Center

Bronx

The introduction of targeted therapies and immunotherapy for early-stage lung cancer is associated with improved survival, but patients can only benefit if they partake in adjuvant and neoadjuvant therapies. Data has shown that inequalities exist for patients with lower socioeconomic status as well as non-White patients when it comes to being referred for and receiving treatment after surgery. These inequalities are likely to increase as new drugs are developed in clinical trials comprised of predominantly white patients. In this project, Dr. Nobel will study the impact of disparities on uptake of adjuvant therapy for NSCLC in a largely minority patient population at Montefiore Medical Center in Bronx, NY. She will provide social support and health literacy to engage patients in their care and collect genetic data about their tumors, which will contribute to future clinical trials that are more inclusive.

Research Summary

Systemic therapy after surgery to remove lung cancer has been demonstrated to improve survival. However, data has shown that there are inequalities in which patients are referred for and receive treatment after surgery, specifically for lower socioeconomic status and non-White patients. As new treatments have been developed, these inequalities are likely to increase as these drugs have been developed in clinical trials predominantly composed of White patients and the benefits in other populations are not known. We have previously demonstrated that using nursing and peer navigators to help guide patients in their cancer care improves treatment adherence in our predominantly Black and Hispanic low socioeconomic status population in the Bronx. The BRONx-TEAM project aims to improve patient outcomes by using a navigation pathway focused on increasing patient adherence to systemic therapy after surgery for non-small cell lung cancer resection. We believe that by providing social support and improving health literacy we can get patients to be more informed and engaged in their cancer care. Furthermore, we will gather genetic data about the patients tumors. Given our patient population, we have a unique opportunity to contribute to the literature to understand the relationships between tumor genetics, treatment types and outcomes in non-White patients. Furthermore, we will investigate the use of a commercial genetic panel to assess risk for recurrence. Given the lack of this type of data in low income non-White patients, we believe that this exploratory portion of our study will serve as an important foundation for future clinical trials that are more inclusive than the currently available literature.

Technical Abstract

As seen in the phase III trial CheckMate 816 (CM816), neoadjuvant anti-PD-1+chemotherapy improves survival for patients with resectable non-small cell lung cancer (NSCLC), with pathologic response as a major trial endpoint. Our team led the Central Pathology Review for CM816, and we showed the first prospective evidence that the full spectrum of % residual viable tumor (%RVT) associates with event free survival. Given the data supporting pathologic response as a survival surrogate, %RVT will likely be incorporated into the next generation of clinical trials and may ultimately guide clinical decision-making. %RVT is primarily evaluated using visual assessment of routine hematoxylin and eosin-stained slides. We developed a machine learning-based approach to score %RVT, which allows for a standardized approach that can be completed rapidly for a large volume of patients, and we propose to test this algorithm in resection specimens from CM816. Additionally, we will use multiplex immunofluorescence (mIF) to quantify individual features of pathologic response, locate them within the larger tumor bed, and determine the relative contribution in predicting patient outcomes. Furthermore, we will use the novel AstroPath platform, a mIF whole-slide imaging platform that uses algorithms first developed in astronomy to generate tumor-immune maps, to identify additional pre- and on-treatment biomarkers of response. Our goal is to leverage emerging technologies (i.e, machine learning and mIF) to develop the next generation of pathology biomarkers, including pathologic response assessment, and to identify additional features that can potentially be targeted in combination with anti-PD-(L)1+chemotherapy to improve clinical benefit in patients with NSCLC.

Key words

Biomarker or biomarker testing

Adjuvant or neoadjuvant

Molecular profile or molecular testing

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Next-generation pathologic response assessment in patients with lung cancer

2024

Career Development Award

Julie Deutsch, MD

Johns Hopkins School of Medicine

Baltimore

Dr. Deutsch’s proposal centers around finding better pathologic predictors of response to neoadjuvant IO in early stage NSCLC. She will utilize machine learning/artificial intelligence to test an algorithm that she and her team have developed that assesses percent residual viable tumor (%RVT), which is the amount of tumor left at the time of surgery. Dr. Deutsch will also characterize tissue specimens using a novel immunofluorescence platform to identify cell types and spatial relationships that are associated with patient benefit to immunotherapy+chemotherapy. This approach can help inform which patients should receive a given therapy, how they will respond, and additional possible targets for the development of new therapies.

Research Summary

Immunotherapy revolutionized the treatment of lung cancer, and is now being extended so patients can receive therapy before surgery. This was supported by a large clinical trial, CheckMate 816 (CM816), where patients with lung cancer showed improved survival when treated with immunotherapy+chemotherapy before surgery, compared to chemotherapy alone followed by surgery. However, there is an unmet need to identify who is most likely to benefit from such an approach. To address this gap, we will apply novel, next-generation pathology biomarkers utilizing machine learning/artificial intelligence and multispectral imaging. Specifically, we have shown that the amount of tumor left at the time of surgery, termed percent residual viable tumor (%RVT), predicts survival. To date, %RVT assessment is primarily performed visually on glass slides using a light microscope. We developed a machine learning-based algorithm for assessing %RVT on digitized glass slides using a small cohort of patients at Johns Hopkins to improve standardization and throughput in preparation for broad usage. Here, we will test the algorithm’s performance in a larger cohort of patients (the CM816 patients). Additionally, we will characterize tissue specimens using the novel multiplex immunofluorescence AstroPath platform, which uses algorithms first developed in astronomy, to identify cell types and spatial relationships that are associated with patient benefit to immunotherapy+chemotherapy. Our goal is to use cutting-edge technologies to improve the care of lung cancer patients by informing which patients should receive a given therapy, how well patients will do after receiving therapy, and possible additional targets for the development of new therapies.

Technical Abstract

Key words

Biomarker or biomarker testing

Adjuvant or neoadjuvant

Molecular profile or molecular testing

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Role of KIRs in Regulating Anti-tumor Immunity and Autoimmunity

2023

Career Development Award

Diane Tseng, MD, PhD

University of Washington and Fred Hutchinson Cancer Center

Seattle

Checkpoint immunotherapy has advanced treatment of NSCLC, but the majority of patients do not experience long-term disease control and are at risk for autoimmune-related side effects. In this study, Dr. Tseng will examine specialized cells called CD8+ T that express receptors (KIR+) that suppress autoimmunity to understand how these cells regulate the immune system’s cancer-fighting ability during checkpoint immunotherapy treatment. Insights gained from this study could result in better strategies for improving efficacy while decreasing immune-related side effects.

Research Summary

Checkpoint immunotherapy has greatly advanced the treatment of non-small cell lung cancer, but only a minority of patients experience long-term disease control and all patients are at risk for side effects related to inflammation in normal organs (known as autoimmunity). Notably, patients developing mild autoimmune side effects are reported to have improved response rates to checkpoint immunotherapy compared to patients that do not develop side effects. However, the scientific basis of this relationship between treatment efficacy and toxicity is not well understood. There has been a recent scientific breakthrough shedding light on a novel role of an immune cell subset involved in controlling autoimmunity. These specialized immune cells are CD8 T cells that express killer cell immunoglobulin-like receptors (KIR+ CD8+ T cells) and function to eliminate other immune cells that drive autoimmune disease. This study is examining the idea that KIR+ CD8+ T cells play an important role in controlling autoimmune side effects from checkpoint immunotherapy, but may also impact how well the immune system fights the cancer. The first goal of this study is to understand how the normal functions of KIR+ CD8+ T cells break down when patients develop autoimmune side effects from checkpoint immunotherapy. The second goal of this study is to understand the extent to which KIR+ CD8+ T cells regulates the immune system’s cancer-fighting capabilities. Better understanding the relationship between desirable immune responses against tumor and undesirable autoimmune toxicities can lead to better strategies for improving the effectiveness of immunotherapy and minimizing side effects.

Technical Abstract

Checkpoint immunotherapy has advanced the treatment paradigm for non-small cell lung cancer (NSCLC), but only some patients respond to treatment and many patients experience immune related adverse events (irAE). Understanding the distinct and overlapping mechanisms regulating anti-tumor immunity and autoimmunity due to checkpoint immunotherapy is critical for developing more effective and less toxic therapies. A subset of CD8+ T cells expressing killer cell immunoglobulin-like receptors (KIR) have recently been shown to suppress autoimmunity without impairing anti-viral immunity by eliminating autoreactive T cells. We reason that KIR+ CD8+ T cells may also play an important role in regulating autoimmunity arising from checkpoint immunotherapy. Furthermore, we reason that KIR+ CD8+ T cells may also regulate the activity of tumor-reactive T cells to varying degrees, since tumor-reactive T cells are a specialized type of autoreactive T cells. This study examines the hypothesis that KIR+ CD8+ T cells dampen autoimmunity and anti-tumor immunity during checkpoint immunotherapy treatment. To test this hypothesis, we will first interrogate the activity of KIR+ CD8+ T cells during the development of irAE after immune checkpoint blockade (specific aim 1). Next, we will evaluate the impact of KIR+ CD8+ T cells on anti-tumor immunity in NSCLC (specific aim 2). This study may shed light on the biological mechanisms regulating autoimmunity and anti-tumor immunity in lung cancer patients undergoing checkpoint immunotherapy. Insights gained from this study may pave the way toward clinical strategies for improving immunotherapy efficacy while minimizing irAEs.

Key words

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Randomized Phase II Trial of Iadademstat with ICI Maintenance in SCLC

2023

Career Development Award

Noura Choudhury, MD

Memorial Sloan Kettering Cancer Center

New York

Small cell lung cancer (SCLC) is difficult to treat, and most patients diagnosed have a poor prognosis. Most patients with SCLC treated with first line chemoimmunotherapy progress within months of immune checkpoint inhibitor (ICI) maintenance therapy. Previous studies in mice have revealed that SCLC treated with iadademstat and maintenance ICI shows enhanced tumor response compared to ICI alone. Dr. Choudhury will conduct a phase II randomized trial investigating this combination in patients with SCLC versus standard of care ICI alone to evaluate progression free survival.

Research Summary

We will conduct a phase II randomized clinical trial investigating the efficacy of combining iadademstat with maintenance ICI versus standard of care ICI maintenance alone for patients with SCLC. We will analyze cell-free DNA in the blood to identify whether treatment with iadademstat and ICI drives conversion to neuroendocrine-low SCLC subtype, which has not been previously demonstrated in a prospective clinical trial. From on-treatment biopsies, we will perform transcriptomic and immunohistochemical analysis to determine whether treatment with iadademstat and ICI induces immunogenicity in SCLC with increased expression of genes in the antigen presentation pathway.

Technical Abstract

Extensive-stage small cell lung cancer (ES-SCLC) has exceptionally poor prognosis, with median survival of one year from diagnosis. Despite neoantigen abundance, SCLC is poorly immunogenic with low expression of major histocompatibility complex class I (MHC-I). As a result, most patients with SCLC treated with first line chemoimmunotherapy progress within months of immune checkpoint inhibitor (ICI) maintenance therapy. Characterization of SCLC by subtypes defined by expression of master transcription factors (TFs) have suggested that ICI-responsive SCLC tumors are composed of neuroendocrine-low (NE-low) subtype with relatively high expression of MHC-I without POU2F3 and with downregulation of ASCL1 and NEUROD1. The hypothesized transcriptional plasticity of SCLC, or subtype switching in response to therapeutic pressures, suggests that leveraging conversion to NE-low subtype with increased immunogenicity may enhance response to ICI. We have shown that inhibition of lysine-specific demethylase 1 (LSD1) with ICI therapy augments tumor regression in ICI-resistant genetically engineered SCLC mouse models, with subsequent upregulation of MHCI and the antigen presentation pathway and suppression of NE gene signatures. With these data supporting its clinical investigation, we propose a phase 2 randomized trial of iadademstat, an oral LSD1 inhibitor, with ICI maintenance in patients with ES-SCLC who have completed initial chemoimmunotherapy. We hypothesize that combination iadademstat and ICI maintenance therapy will improve progression-free survival and promote conversion to NE-low, immunogenic SCLC. We will trace key SCLC TF activity throughout treatment using a novel cell-free DNA (cfDNA) nucleosome profiling assay and utilize optional on-treatment biopsies to characterize differential gene expression with treatment.

Key words

Small cell lung cancer (SCLC)

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Development of ALK-specific TCR-T cells for the eradication of ALK+ NSCLC

2022

Partner Awards

Grant title (if any)

ALK Positive/LUNGevity Lung Cancer Research Awards

Roberto Chiarle, MD

Boston Children’s Hospital/Harvard Medical School

Boston

In this project, Dr. Chiarle and his team will generate T cells that have engineered receptors, called TCR receptors (TCR-T cells), that will selectively target and attack the ALK protein that is expressed by tumor cells. Generation of such cells could be a powerful tool to eradicate ALK+ lung cancer cells and form the basis of a TCR-T cell-based clinical trial for patients with TKI-resistant ALK+ NSCLC.

Research Summary

Patients with an ALK-positive lung cancer are typically young. For these patients several oral ALK inhibitors are available as pills, typically alectinib, lorlatinib, and brigatinib. These ALK inhibitors are very effective and well tolerated and most patients respond for long time. However, tumors may progressively develop resistance and start to grow back. In this setting of relapsing disease, few therapeutic options are left because most patients with ALK-positive lung cancer do not respond to existing immunotherapies.

For several years, we have been working to develop immunotherapy tools specifically for patients with ALK-positive lung cancer. In this proposal, the strategy is to re-engineer patients’ naturally occurring immune cells to target ALK selectively expressed by tumor cells. First, we will discover several receptors (TCRs) that T lymphocytes use to recognize ALK and kill tumor cells. Next, we will re-introduce these TCRs into the T lymphocytes to rapidly generate a large pool of killer cells all directed simultaneous against the tumor. The rapid infusion of a large number of TCR-engineered T lymphocytes, called TCR-T cells, will generate a wave of killing toward which the tumor will not have the time to adapt and escape. While the identification of ALK specific TCR might be difficult and elusive, we already discovered a series of very specific TCRs directed against ALK expressed by ALK-positive tumors in mice. Leveraging the discovery of ALK peptides expressed by human tumor cells, we will now discover specific TCRs that recognize ALK in human lung cancer. We will use these TCR-T cells to cure ALK-positive human tumors in combination with ALK inhibitors. We expect that this combination will be extremely powerful to rapidly eradicate tumor cells in the lung as well as in metastatic sites.

Technical Summary

About 5-7% of non-small cell lung cancers (NSCLCs) have a rearrangement of the anaplastic lymphoma kinase (ALK) gene. ALK-rearranged NSCLC is typically treated with ALK tyrosine kinase inhibitors (TKIs), but no effective immunotherapies are available for refractory or relapsed tumors. In previous work, we have shown that ALK is an immunogenic oncoprotein that induces specific T cell responses with potent killing activity against ALK+ tumor cells in the lung and in distant metastatic sites, such as the brain. Thus, the lack of efficacy of standard immunotherapy in ALK+ NSCLC can be bypassed by the development of tools to induce ALK-specific T cell responses. Our central hypothesis is that the generation of T cells engineered to express TCRs (TCR-T cells) that selectively target ALK+ NSCLC will be a powerful tool to completely eradicate ALK+ lung cancer cells.

In preliminary experiments, we have shown the feasibility of the cloning of ALK-specific TCRs against an ALK peptide expressed by a mouse model of ALK+ lung cancer. In Aim 1, we will test the potency, the safety and the in vivo efficacy of ALK-specific TCR-T cells obtained by T cells engineered to express TCRs against ALK in a mouse model of ALK+ lung cancer. TCR-T cells will be tested alone or in combination with ALK TKI. Their efficacy will be compared to natural anti-ALK T cells induced by vaccination in mice. In Aim 2, we will discover and study the activity of ALK-specific TCRs against human ALK peptides. We will leverage ALK peptides we previously identified to be expressed by human ALK+ NSCLC. The killing activity of these human specific TCR-T cells will be tested against human ALK+ NSCLC in vitro and in vivo, alone or in combination with ALK TKIs. We expect to identify potent and specific TCRs as leading candidates for a TCR-T cell-based clinical trial for patients with TKI-resistant ALK+ NSCLC.

Key words

Acquired resistance

Anaplastic lymphoma kinase (ALK)

Targeted therapy

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Tumor draining lymph node immunomodulation to decrease recurrence in NSCLC

2022

Health Equity and Inclusiveness Junior Investigator Award

Jonathan Villena-Vargas, MD

Weill Medical College of Cornell University

New York

Lymph nodes are small structures that work as filters for foreign substances, such as cancer cells and infections. These nodes contain infection-fighting immune cells that are carried in through the lymph fluid. This project will study the lymph node draining basin, which is involved in the spread of a tumor from the original location site to distant sites, and whether activating cancer-fighting T-cells can decrease recurrence in NSCLC. Dr. Villena-Vargas will use animal models to investigate whether immune checkpoint inhibitors enhance lymph node T-cells memory, which increases their ability to recognize cancer cells in the bod and can prevent metastatic recurrence.

Research Summary

Approximately 25% of the patients diagnosed with non-small cell lung cancer (NSCLC) exhibit early-stage disease. Patients with early-stage (I/II) NSCLC are routinely treated by surgery, which results in local control. However, systemic relapse (metastasis) occurs in 50% patients within 5 years of surgical intervention even with additional therapy such as chemotherapy and/or radiation. Immune checkpoint inhibitors (ICI) targeting the PD-1/PD-L1 axis have revolutionized the treatment landscape, generating therapeutic efficacy in approximately 25% of patients in early and advanced stages of lung cancer. However, there are currently inconclusive studies as to 1) timing of treatment 2) optimal combination therapy 3) prediction of patient response. In order, to recapitulate early-stage lung cancer treatment and metastatic recurrence, we have established a murine model of resection and recurrence. Our preliminary data has revealed a major immune response at the level of the tumor draining lymph node (TDLN) with PD-1 checkpoint blockade. In addition, this response is primarily responsible for T-cell memory differentiation, proliferation, and survival with subsequent decrease in metastatic recurrence in responding mice. Given these results, we hypothesize that optimal therapy against metastatic recurrence will be dependent on establishment of appropriate T-cell memory “immunosurveillance”. In addition, we have shown that the memory nodal response can be augmented by utilizing a cytokine immunomodulatory combination strategy with cure in over 70% of mice. Importantly, we have characterized that these specific T-cell memory population are uniquely found in the TDLN of early-stage patients (i.e. not in the primary tumor or peripheral blood that is commonly interrogated). In this proposal we will utilize a combination of both novel preclinical mouse models and patient-derived tumor draining lymph node T cells, to dissect molecular and cellular mechanisms of “immune-memory” response or resistance to immunotherapy, the key steps in establishing immunosurveillance for freedom from metastatic recurrence.

Technical Abstract

Anti–PD-1 therapy has transformed the management of NSCLC, but only a subset of patients respond to anti–PD-1 therapy, and responses are rarely curative. Understanding the effects of anti–PD-1 therapy on the composition and functional state of tumor-associated immune subsets can identify mechanisms of response and resistance to anti–PD-1 therapy and provide insights into rational combination strategies to improve upon response rates. Although the effects of anti–PD-1 therapy within the tumor immune microenvironment in preclinical and human subjects have been previously reported, we propose to broadly characterize the effects of anti–PD-1 therapy on the tumor draining lymph nodes (TDLN). Studying the TDLN is particularly valuable because it is the primary sites at which the tumor antigen exposure and the main site of immune T-cell differentiation. To determine the effect of ICI response in the lymph node, we have created an immunocompetent murine model that recapitulates early-stage treatment and outcomes in patients. Additionally, we have characterized TDLN T cells from early-stage cancer patients and have been able to recapitulate our preclinical findings of unique memory T-cell subsets. Our preliminary results of the immune cell type composition, checkpoint expression, and cytokine expression within TDLNs suggest that there are major remodeling events within the TDLN mediated by anti–PD-1 therapy that are critical for conferring effective antitumor long-term immune responses. We demonstrate that unique memory T cell subsets undergo significant activation, expansion, and maturation in response to anti–PD-1 therapy, to shape antitumor programming in TDLNs. Importantly, our proposed study will provide a methodological demonstration in both a murine model and human TDLN that simultaneous uses Flowcytometry, functional immune assays and single cell sequencing that will enable deep interrogation of the immune profile to PD-1 inhibition resistant and responsive patients.

Key words

Squamous cell lung cancer