Cynthia Zhao

All Publications

• Rewiring oncogenic signalling to precision ablation of metastatic cancer. Nature biomedical engineering Zou, X., Palafox, E., Zhao, C., Beier, K. T., Kang, C. Y., Lin, M. Z. 2026

  Abstract

  Despite recent advances, long-term survival in metastatic carcinomas such as ovarian cancer remains limited by off-tumour toxicities of targeted therapies and low response rates to immunotherapy. Synthetic proteins have been engineered for selective recognition of oncogenic signalling states, but how they can be used to treat metastatic disease in vivo remains unclear. Addressing cancers driven by ErbB-family receptor tyrosine kinases such as EGFR and HER2, we used engineered proteins to restrict replication of a clinically approved viral backbone to kill cells with aberrant ErbB signalling. The resulting ErbB oncogene-selective virus (ErbB-OSV) showed superior safety to a benchmark oncolytic virus of the same family and superior efficacy against ErbB2/HER2-positive ovarian cancer xenografts. In a syngeneic model of advanced ovarian cancer, combining ErbB-OSV with chemotherapy and enabling repeated dosing by B cell depletion conferred a 180% larger survival benefit compared to chemotherapy alone, while single-agent ErbB-OSV cured most early cases. Thus, rationally restricting viral replication to ErbB-hyperactive cells with synthetic signalling proteins yields a highly specific therapeutic agent that ablates metastatic tumours in vivo more effectively than existing treatments.

  View details for DOI 10.1038/s41551-026-01704-9

  View details for PubMedID 42286252

  View details for PubMedCentralID 7067809

• In vivo CRISPR screening in head and neck cancer reveals Uchl5 as an immunotherapy target. Nature communications Fu, C., Saddawi-Konefka, R., Chinai, J. M., Kim, S. Y., Kammula, A. V., Perera, J. J., Jiang, A., Tiwari, P., Kistler, E. N., Tang, S., Luna, S. M., Colvin, K. J., Dubrot, J., Anderson, S., Fetterman, R. A., Chuong, C. L., Lane-Reticker, S. K., Cheruiyot, C. K., Muscato, A. J., Alipour, Z., Adkins, D. R., Griffin, G. K., Bernstein, B. E., Egloff, A. M., Yates, K. B., Chernock, R. D., Gutkind, J. S., Uppaluri, R., Manguso, R. T. 2025; 16 (1): 8572

  Abstract

  Recurrent/metastatic head and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy with a significant unmet need for enhancing immunotherapy response given current modest efficacy. Here, we perform an in vivo CRISPR screen in an HNSCC mouse model to identify immune evasion genes. We identify several regulators of immune checkpoint blockade (ICB) response, including the ubiquitin C-terminal hydrolase 5 (UCHL5). Loss of Uchl5 in tumors increases CD8+ T cell infiltration and improved ICB responses. Uchl5 deficiency attenuates extracellular matrix (ECM) production and epithelial-mesenchymal-transition (EMT) transcriptional programs, which contribute to stromal desmoplasia, a histologic finding we describe as associated with reduced anti-PD1 response in human HNSCCs. COL17A1, a collagen highly and specifically expressed in HNSCC, mediates in part Uchl5-mediated immune evasion. Our findings suggest an unappreciated role for UCHL5 in promoting EMT in HNSCC and highlight ECM modulation as a strategy to improve immunotherapy responses.

  View details for DOI 10.1038/s41467-025-63592-y

  View details for PubMedID 41022734

  View details for PubMedCentralID PMC12480505

• Longitudinal liquid biopsy identifies an early predictive biomarker of immune checkpoint blockade response in head and neck squamous cell carcinoma. Nature communications Wang, B., Saddawi-Konefka, R., Clubb, L. M., Tang, S., Wu, D., Mukherjee, S., Sahni, S., Dhruba, S. R., Yang, X., Patiyal, S., Day, C. P., Desai, P. A., Allen, C., Wang, K., Gutkind, J. S., Ruppin, E. 2025; 16 (1): 8161

  Abstract

  Immune checkpoint blockade (ICB) has improved outcomes for patients with head and neck squamous cell carcinoma (HNSCC), but predictive biomarkers remain limited. Here, we use a time-resolved, multi-omic approach in a murine HNSCC model to characterize peripheral immune responses to ICB. Single-cell transcriptomics and T/B cell receptor analyses  reveal early on-treatment expansion of effector memory T and B cell repertoires in responders, preceding tumor regression. These dynamic immune features inform a composite transcriptional signature that accurately predicts ICB response in independent human HNSCC cohorts. LiBIO outperforms existing biomarkers and generalizes to melanoma, non-small cell lung cancer, and breast cancer without retraining. These findings suggest that early treatment-induced changes in circulating immune repertoires reflect the host's capacity to mount an effective antitumor response. This work provides a framework for leveraging transient peripheral immune dynamics to develop non-invasive, high-fidelity biomarkers for response to immunotherapy across cancer types.

  View details for DOI 10.1038/s41467-025-63538-4

  View details for PubMedID 40890155

  View details for PubMedCentralID PMC12402333

• The tumor-sentinel lymph node immuno-migratome reveals CCR7<SUP>+</SUP> dendritic cells drive response to sequenced immunoradiotherapy NATURE COMMUNICATIONS Saddawi-Konefka, R., Al Msari, R., Tang, S., Philips, C., Sadat, S., Clubb, L. M., Luna, S., Fassardi, S., Jones, R., Pietryga, I., Faraji, F., Schokrpur, S., Yung, B. S., Allevato, M. M., Decker, K. E., Nasamran, C. A., Chilin-Fuentes, D., Rosenthal, S., Jensen, S. M., Fox, B. A., Bell, R., Gutkind, J., Sharabi, A., Califano, J. A. 2025; 16 (1): 6578

  Abstract

  Surgical ablation or broad radiation of tumor-draining lymph nodes can eliminate the primary tumor response to immunotherapy, highlighting the crucial role of these nodes in mediating the primary tumor response. Here, we show that immunoradiotherapy efficacy is dependent on treatment sequence and migration of modulated dendritic cells from tumor to sentinel lymph nodes. Using a tamoxifen-inducible reporter paired with CITE-sequencing in a murine model of oral cancer, we comprehensively characterize tumor immune cellular migration through lymphatic channels to sentinel lymph nodes at single-cell resolution, revealing a unique immunologic niche defined by distinct cellular phenotypic and transcriptional profiles. Through a structured approach of sequential immunomodulatory radiotherapy and checkpoint inhibition, we show that sequenced, lymphatic-sparing, tumor-directed radiotherapy followed by PD-1 inhibition achieves complete and durable tumor responses. Mechanistically, this treatment approach enhances migration of activated CCR7+ dendritic cell surveillance across the tumor-sentinel lymph node axis, revealing a shift from their canonical role in promoting tolerance to driving antitumor immunity. Overall, this work supports rationally sequencing immune-sensitizing, lymphatic-preserving, tumor-directed radiotherapy followed by immune checkpoint inhibition to optimize tumor response to immunoradiotherapy by driving activated dendritic cells to draining sentinel lymph nodes.

  View details for DOI 10.1038/s41467-025-61780-4

  View details for Web of Science ID 001531633000005

  View details for PubMedID 40675962

  View details for PubMedCentralID PMC12271439

• CHMP2A regulates broad immune cell-mediated antitumor activity in an immunocompetent in vivo head and neck squamous cell carcinoma model JOURNAL FOR IMMUNOTHERAPY OF CANCER Yun, J., Saddawi-Konefka, R., Goldenson, B., Al-Msari, R., Bernareggi, D., Thangaraj, J. L., Tang, S., Patel, S. H., Luna, S. M., Gutkind, J., Kaufman, D. 2024; 12 (5)

  Abstract

  Natural killer (NK) cells are key effector cells of antitumor immunity. However, tumors can acquire resistance programs to escape NK cell-mediated immunosurveillance. Identifying mechanisms that mediate this resistance enables us to define approaches to improve immune-mediate antitumor activity. In previous studies from our group, a genome-wide CRISPR-Cas9 screen identified Charged Multivesicular Body Protein 2A (CHMP2A) as a novel mechanism that mediates tumor intrinsic resistance to NK cell activity.Here, we use an immunocompetent mouse model to demonstrate that CHMP2A serves as a targetable regulator of not only NK cell-mediated immunity but also other immune cell populations. Using the recently characterized murine 4MOSC model system, a syngeneic, tobacco-signature murine head and neck squamous cell carcinoma model, we deleted mCHMP2A using CRISPR/Cas9-mediated knock-out (KO), following orthotopic transplantation into immunocompetent hosts.We found that mCHMP2A KO in 4MOSC1 cells leads to more potent NK-mediated tumor cell killing in vitro in these tumor cells. Moreover, following orthotopic transplantation, KO of mCHMP2A in 4MOSC1 cells, but not the more immune-resistant 4MOSC2 cells enables both T cells and NK cells to better mediate antitumor activity compared with wild type (WT) tumors. However, there was no difference in tumor development between WT and mCHMP2A KO 4MOSC1 or 4MOSC2 tumors when implanted in immunodeficient mice. Mechanistically, we find that mCHMP2A KO 4MOSC1 tumors transplanted into the immunocompetent mice had significantly increased CD4+T cells, CD8+T cells. NK cell, as well as fewer myeloid-derived suppressor cells (MDSC).Together, these studies demonstrate that CHMP2A is a targetable inhibitor of cellular antitumor immunity.

  View details for DOI 10.1136/jitc-2023-007187

  View details for Web of Science ID 001222050100002

  View details for PubMedID 38702144

  View details for PubMedCentralID PMC11086353