Jos Melenhorst Laboratory

Research

Overview

The Melenhorst lab is interested in understanding and improving tumor immunity. Our translational T cell and tumor immunology research focuses on the use of chimeric antigen receptor (CAR)-engineered T cells that are specific to antigens expressed by tumor cells. We use basic T cell immunology research as a starting point to study the interactions between CAR T cells and tumor cells, as well as those between CAR T cells and tumor microenvironments. We also study patient responses to CAR T-cell therapies using correlative, biomarker, and mechanistic approaches. These have led to the discovery of general themes of response and resistance to these therapies in various hematologic malignancies and the epigenetic and metabolomic underpinnings of CAR T-cell efficacy. We use high-throughput genomic and pharmacological methods to interrogate the various pathways as well as novel PDX and syngeneic preclinical models of human cancers. The ultimate aim of our lab is to develop safe and effective cell-based therapies for cancer. Knowing what defines efficacy is our first goal. The second is determining how we can improve upon such efficacy rates.

Several major efforts in our lab include:
1. Linking insertional mutagenesis and cell function to improve CAR-T cell therapy:

Chimeric antigen receptors can be delivered to T cells through viral or nonviral vectors. Lentiviral vectors integrate their genetic content into the host cell’s DNA in a relatively random way, targeting both intra- and intergenic regions. Over the course of several years, these vectors have been optimized for safety and efficiency, and FDA-approved lentiviral-based CAR T cells have shown remarkable efficacy against B-cell malignancies. In a previous mechanistic study performed by our group, we observed that insertional mutagenesis in the TET2 locus was associated with complete response (CR) and enhanced CAR T cell function in a Chronic Lymphocytic Leukemia (CLL) patient. A follow-up study applying integration site analysis in a bigger cohort – including subjects with acute lymphoblastic leukemia (ALL) and CLL – revealed that it was possible to associate insertion sites with the outcomes presented by the patients: CR, partial response with transformed disease, or no response. Based on these findings, we optimized a pipeline to screen for genes that are disrupted by CAR insertions; we hope to identify proteins and pathways associated with superior CAR T-cell proliferation and persistence, which, therefore, will lead to the development of a more effective product. 

2. Improving CAR T-cell manufacturing to achieve long-term remissions in hematological malignancies

CAR T-cell therapy can induce complete remissions in B cell leukemias and lymphoma (anti-CD19 CAR T cells), as well as multiple myeloma (anti-BCMA CAR T cells), but a significant proportion of patients will ultimately relapse. Our goal is to improve CAR T-cell manufacturing to increase long-term persistence and remission rates, while limiting toxicities and interindividual variabilities, and to reduce the duration and costs of manufacturing. This is achieved through a comprehensive assessment of patient-derived CAR T cells to identify mechanisms of therapeutic success or failure. Previous translational studies have identified several factors as determinants of remission and durable response to CD19 CAR T-cell therapy of CLL, including higher frequency of early memory T cells and IL-6/STAT3-pathway upregulation (PMID: 29713085). Therefore, our goal is to improve the manufacturing process to enrich the final product with those cells responsible for the efficiency. This is achieved by selecting only these lymphocytes for the manufacturing process and by optimizing the culture conditions to promote the preservation or the emergence of these characteristics. 

3. Developing CAR T cell therapy for acute myeloid leukemia 

Contrary to the breakthrough of CAR T cells in lymphoid hemopathies, the treatment of myeloid neoplasm – including acute myeloid leukemia (AML) and high-risk myelodysplastic syndrome (MDS) –has not yet been revolutionized by CAR T cells. The prognosis for these diseases remains poor, and the only curative treatments – including heavy chemotherapy and allogeneic hematopoietic stem cell transplantation – are aggressive with limiting toxicities (high morbidity and mortality rates). The difficulties in developing anti-AML CAR T cells are due in part to the greater heterogeneity of this disease, with the absence of an obvious antigenic target expressed on the surface of all leukemic blasts and not expressed on the surface of healthy cells. Our goal is to study the expression of certain antigens of interest in AML subtypes, and to develop CAR T cells to treat these specific subtypes. Furthermore, given the importance of lymphocyte fitness for CAR T-cell function, we also study the phenotypic and functional alterations of lymphocytes from AML patients both at diagnosis and after chemotherapy in order to further optimize the manufacturing of these CAR T cells. 

4. Epigenetic, metabolomic engineering of CAR T cells 

Chronic lymphocytic leukemia (CLL) exhibits a poor clinical response to current CAR T-cell therapy, making it one of the most challenging forms of leukemia. In 2018, our group reported a case with complete remission of CLL upon CAR T-cell therapy and discovered clonal expansion of CD8+CAR T cells with hypomorphic disruption on TET2 locus by lentiviral integration of CD19BBz-CAR (CAR19). We investigated the idea that clonal expansion in CAR T cells was highly correlated to early memory function improvement. We have expanded our knowledge of TET2 in CAR19 T cells into functional disruption on a post-translational stage.

1. Accumulation of D-2-hydroxyglutarate by Isocitrate dehydrogenases 1 (IDH1) neomorph overexpression in CAR19 T cells. D-2-HG works as weak antagonist to multiple a-KG dependent dioxygenase, including TET family of 5-methylcytosine hydroxylases. We hypothesized that D-2-HG accumulation by IDH1 neomorph overexpression in CAR19 T cells leads into improvement of proliferation and memory function by functional disruption of TET2.

In the News

Films
Fire With Fire | Directed by Ross Kauffman, Vimeo

Of Medicine and Miracles | Directed by Ross Kauffman, Tribeca Film Festival

Articles
T-cell immunotherapy tied to 10-year remission in two leukemia patients, study finds | CNN

Leukaemia-fighting CAR T-cells kept patients in remission for 10 years - and counting | ABC Science

How CAR T-Cell Therapy 'Cured' 2 Cancer Patients | Verywell Health

Researchers label early CAR-T therapy patient 'cured' after living a decade without cancer | STAT

Single-Cell Multi-Omics Reveals Type-2 Fuctionality in CAR T Cell | Oncology Tube ASH 2023

Podcasts
Future of CAR T-Cell Therapy | Cancer Advances Podcast

Optimizing CAR T-Cell and Stem Cell Transplants | Cancer Advances Podcast

Selected Publications

View publications for Jos Melenhorst, PhD
(Disclaimer: This search is powered by PubMed, a service of the U.S. National Library of Medicine. PubMed is a third-party website with no affiliation with Cleveland Clinic.)

Zhao, Z., & Melenhorst, J. J. (2022). Adaptable Leukemia Cells Resisting CAR T-cell Attack via B-cell Activation. Cancer immunology research, 10(9), 1040. 

J. Joseph Melenhorst, Gregory M. Chen, Meng Wang, David L. Porter, Changya Chen, McKensie A. Collins, Peng Gao, Shovik Bandyopadhyay, Hongxing Sun, Ziran Zhao, Stefan Lundh, Iulian Pruteanu-Malinici, Christopher L. Nobles, Sayantan Maji, Noelle V. Frey, Saar I. Gill, Lifeng Tian, Irina Kulikovskaya, Minnal Gupta, David E. Ambrose, Megan M. Davis, Joseph A. Fraietta, Jennifer L. Brogdon, Regina M. Young, Anne Chew, Bruce L. Levine, Donald L. Siegel, Cécile Alanio, E. John Wherry, Frederic D. Bushman, Simon F. Lacey, Kai Tan, Carl H. June: Decade-long leukaemia remissions with persistence of CD4+ CAR T cells. Nature (in press) 2022.   

Kavita M. Dhodapkar, Adam D. Cohen, Akhilesh Kaushal, Alfred L. Garfall,   Renee Julia Manalo, Allison R. Carr,  Samuel S. McCachren, III, Edward A. Stadtmauer, Simon F. Lacey, J. Joseph Melenhorst, Carl H. June, Michael C. Milone, Madhav V. Dhodapkar: Changes in Bone Marrow Tumor and Immune Cells Correlate with Durability of Remissions Following BCMA CAR T Therapy in Myeloma." Blood Cancer Discovery  3(6): 490-501, 2022.

Arya, Rahul, Barrett, David M.,  Grupp, Stephan A., Melenhorst, Jan Joseph:  Improving and Maintaining Responses in Pediatric B–Cell Acute Lymphoblastic Leukemia Chimeric Antigen Receptor–T Cell Therapy." The Cancer Journal 27(2). 2021.

Nobles CL, Sherrill-Mix S, Everett JK, Reddy S, Fraietta JA, Porter DL, Frey N, Gill SI, Grupp SA, Maude SL, Siegel DL, Levine BL, June CH, Lacey SL, Melenhorst JJ+, Bushman FD+: CD19-targeting CAR T cell immunotherapy outcomes correlate with genomic modification by vector integration. Journal of Clinical Investigation 130(2): 673-685, 2020. 

Stadtmauer, EA, Fraietta, JA, Davis, MM, Cohen, AD, Weber, KL, Lancaster, E, Mangan, PA, Kulikovskaya, I, Gupta, M, Chen, F, Tian, L, Gonzalez, VE, Xu, J, Jung, IY, Melenhorst, JJ, Plesa, G, Shea, J, Matlawski, T, Cervini, A, Gaymon, AL, Desjardins, S, Lamontagne, A, Salas-Mckee, J, Fesnak, A, Siegel, DL, Levine, BL, Jadlowsky, JK, Young, RM, Chew, A, Hwang, WT, Hexner, EO, Carreno, BM, Nobles, CL, Bushman, FD, Parker, KR, Qi, Y, Satpathy, AT, Chang, HY, Zhao, Y, Lacey, SF, June CH: CRISPR-engineered T cells in patients with refractory cancer. Science 367(6481): eaba7365, Feb 2020.

Van Bruggen JAC, Martens AWJ, Fraietta JA, Hofland T, Tonino SH, Eldering E, Levin MD, Siska PJ, Endstra S, Rathmell JC, June CH, Porter DL, Melenhorst JJ, Kater AP, van der Windt GJW: Chronic lymphocytic leukemia cells impair mitochondrial fitness in CD8+ T cells and impede CAR T cell efficacy. Blood Page: pii: blood.2018885863, May 2019. 

Cohen AD, Garfall AL, Stadtmauer EA, Melenhorst JJ, Lacey SF, Lancaster E, Vogl DT, Weiss BM, Dengel K, Nelson A, Plesa G, Chen F, Davis MM, Hwang WT, Young RM, Brogdon JL, Isaacs R, Pruteanu-Malinici I, Siegel DL, Levine BL, June CH, Milone MC.: B cell maturation antigen-specific CAR T cells are clinically active in multiple myeloma. The Journal of Clinical Investigation 130(6): 2210-2221, Mar 2019.

Ruella M, Xu J, Barrett DM, Fraietta JA,Reich T, Ambrose DE, Klichinsky M, Shestova O,Patel PR, Kulikovskaya I, Nazimuddin F, Bhoj VG, Orlando E,Bitter H, Maude SL, Levine BL, Nobles CL,Bushman FD, Young RM, Scholler J, Gill SI,  Fry TJ, June CH, Grupp SA, Lacey SF, Melenhorst JJ: Induction of Resistance to Chimeric Antigen Receptor (CAR) T Cell Therapyby Transduction of a Single Leukemic B Cell. Nature Medicine 24(10): 1499-1503, October 2018. 

Joseph A. Fraietta, Christopher L. Nobles, Morgan A. Sammons, Stefan Lundh, Shannon A. Carty, Tyler Reich, Alexandria P. Cogdill, Jennifer J.D. Morrissette, Jamie E. DeNizio, Shantan Reddy, Young Hwang, Mercy Gohil, Irina Kulikovskaya, Farzana Nazimuddin, Minnal Gupta, Fang Chen, John K. Everett, Katherine A. Alexander, Enrique Lin-Shiao, Marvin H. Gee, Xiaojun Liu, Regina M. Young, David Ambrose, Yan Wang, Jun Xu, Martha S. Jordan, Katherine T. Marcucci, Bruce L. Levine, K. Christopher Garcia, Yangbing Zhao, Michael Kalos, David L. Porter, Rahul M. Kohli, Simon F. Lacey, Shelley L. Berger, Frederic D. Bushman, Carl H. June, and J. Joseph Melenhorst: Disruption of TET2 Promotes the Therapeutic Efficacy of CD19-targeted T-cells. Nature 558(7709): 307-312, June 2018.