Ruella Research

At the Ruella Lab, we are dedicated to transforming the treatment of lymphoid malignancies through cutting-edge cellular immunotherapy. Our research spans the full spectrum from basic science to translational discovery to early-phase clinical trials, with a singular mission: to improve outcomes for patients with lymphoma, leukemia, myeloma and cancer.

A hallmark of our program is its deeply translational nature. We systematically biobank tumor and blood samples from patients treated with commercial CAR T-cell therapies and bispecific antibodies, enabling us to study real-world responses, resistance mechanisms, and toxicities at an unprecedented level of detail. This rich clinical resource fuels our basic and preclinical research and drives the design of next-generation therapies.

We have made pioneering contributions to the understanding of how tumors escape CAR T-cell therapy, including the first report of a leukemic cell acquiring a CAR gene, the discovery of apoptosis pathway defects conferring resistance, and the role of the gut microbiome in influencing treatment efficacy. We have also developed innovative solutions—such as engineered CARs that resist venetoclax-induced apoptosis or target novel CD19 epitopes—and brought these concepts into early human trials.

Our current research continues to break new ground. We are investigating CD5 and BTLA as novel immune checkpoints, developing CAR T cells that selectively target malignant clones while sparing normal B cells, and exploring how diet, antibiotics, and the epigenome can modulate immune responses. Our preclinical models—including animal and patient-derived systems—are central to these efforts and have led to the translation of multiple therapeutic concepts to the clinic.

Through collaborations with clinicians and scientists at Penn and beyond, we harness the power of next-generation sequencing, proteomics, metabolomics, and spatial transcriptomics to uncover the complex interplay between tumors and the immune system.

Ultimately, the Ruella Lab strives to transform scientific insights into curative therapies. Our work has not only shaped the field of immunotherapy but continues to redefine what is possible for patients with otherwise intractable blood cancers.

 

Here is an overview of our past and current research:

1. Resistance Mechanisms to CAR T-Cell Immunotherapy

• The role of apoptosis in both tumor and CAR T cells: In 2022, we showed how BCL-2 orchestrates responses to CART19 in lymphoma and demonstrated that targeting BCL-2 in both tumor and T cells enhances efficacy (Lee YG, Cancer Discov, 2022). Previously, we identified the extrinsic apoptosis—or “death receptor”—pathway as a critical node for CART19 resistance in B-ALL, confirmed by two clinical trials (Singh N, Cancer Discov, 2020).
• The role of the lymphoma tumor microenvironment and the gut microbiota: We were the first to demonstrate the impact of the gut microbiota on CAR T-cell immunotherapy, showing that specific taxa within the class Clostridia correlate with response to therapy (Smith M, Nat Med, 2022; Uribe-Herranz M, Mol Ther, 2023) (WO2023081472A1). We also defined the role of the BTLA:HVEM axis in the tumor microenvironment and engineered BTLA knockout CAR T cells with enhanced efficacy (Guruprasad P, Nat Immunol, 2024). Earlier, we showed that CAR T cells targeting CD123 could kill Hodgkin lymphoma cells and reprogram tumor-associated macrophages (TAMs), enhancing therapeutic outcomes (Ruella M, Cancer Discov, 2017) (US2019161542A1).
• Mechanism of antigen-negative escape: We described for the first time a case of relapse due to aberrant expression of the anti-CD19 CAR by leukemic blasts after accidental transduction during manufacturing. This discovery led to changes in CAR T-cell production protocols (Ruella M, Nat Med, 2018) (WO2019089798A1).

2. Development of Next-Generation CAR T-Cell Therapies for Cancer and Autoimmune Disease

• CAR T cells for T-cell malignancies: We developed a novel CD5-deleted anti-CD5 CAR T-cell therapy with enhanced efficacy (Patel RP, Sci Immunol, 2024), leading to the formation of Vittoria Biotherapeutics and a phase I trial (Senza5 CART5). We are also advancing anti-CD2 CAR T-cell therapies (Angelos M, ASH 2023, #885).
• Targeting B-cell neoplasms and autoimmune diseases: We created a platform to selectively target IGHV and IGLV sequences enriched in B-cell lymphomas, leukemias, and autoimmune diseases. Our anti-IGHV4-34 CAR T-cell product is in preclinical testing for lymphoma and lupus (Cohen I, ASH 2023 #1020) (WO2023023596A1).
• Enhancing CAR T efficacy and safety with small molecules: We showed that the BTK inhibitor ibrutinib enhances anti-lymphoma CAR T-cell activity (Ruella M, Clin Cancer Res, 2016) (US2018140602A1; WO2018013918A3), a concept validated in clinical trials (Gill S, Blood Adv, 2022). More recently, we demonstrated that SMAC mimetics and BCL-2 inhibitors synergistically boost CAR T-cell killing (Lee YG, Cancer Discov, 2022) (WO2019165215A1; WO2023019165A1).
• Targeting CD19-negative relapses: We were the first to demonstrate that dual targeting of CD19 and CD123 can prevent antigen-loss relapses in leukemia (Ruella M, J Clin Invest, 2016) (US2016028896A1). We also developed CD22 CAR T cells now in clinical trials (Singh N, Nat Med, 2021) (US2023220090A1), and CD79b-targeted CARs (Ghilardi G, ASH 2022). Recently, we published a first-in-human study of a novel CAR19 construct targeting a membrane-proximal CD19 epitope (Zhang Y, Mol Cancer, 2023).

3. CAR T-Cell Therapy–Related Toxicities

• Secondary primary malignancies (SPMs): In response to an FDA safety alert, we were the first to characterize the incidence and nature of SPMs, including CAR-negative T-cell lymphomas, in patients treated with commercial CAR T-cell therapies (Ghilardi G, Nat Med, 2024).
• Bendamustine as safer lymphodepletion: We led four multicenter retrospective studies showing that bendamustine is as effective as traditional fludarabine/cyclophosphamide conditioning but results in fewer toxicities (Ghilardi G, Ann Oncol, 2022; Ghilardi G, Blood Adv, 2024).
• Modulating cytokine release syndrome (CRS): We showed that ibrutinib and ruxolitinib significantly reduce CRS in xenograft models of leukemia and lymphoma treated with CAR T cells (Ruella M, Leukemia, 2017). We are now investigating new targets that are associated with effector T-cell toxicities using high throughput technologies to analyze patient samples.