Research

My long-term research goal is to develop a new combination treatment for glioblastoma that will overcome a highly heterogenous and immunologically cold landscape. My approach for accomplishing this goal will leverage a highly potent new drug developed at Duke – a brain bi-specific T cell engager (BRiTE). BRiTEs can redirect T cells against tumor by targeting tumor-specific or associated antigens, and induce tumor kill at picomolar concentrations. My research focuses on extending and enhancing this strategy by co-administering multiple BRiTEs targeting different tumor antigens. In addition, I propose pre-administration of ex vivo expanded autologous T cells (autologous lymphocyte transfer (ALT)) to inflame the immunologically cold tumor before drug infusion. Together, my aim is to develop an off-the-shelf approach to targeting multiple antigens, while also ensuring the presence of functional immune cells in tumor. With this platform, a library of antigen-targeting BRiTEs can be repeatedly drawn upon and co-administered with ALT, thereby achieving robust and durable control of aggressive cancers like glioblastoma.

This proposed approach builds upon my pre-clinical doctoral research in Biomedical Engineering (immunoengineering specialization), which I began under Dr. John H. Sampson in the fall of 2021 at Duke University. Here I evaluated whether pre-administration of autologous lymphocytes could increase the presence of T cells in established brain tumors. I found that lymphocytes cultured with IL-7 could accumulate in intracranial tumors, even during evidence of endogenous T cell sequestration. Pre-administration of this autologous lymphocyte product also enhanced the efficacy of multiple T cell activating therapies, including BRiTE, across multiple models of murine and patient-derived glioma. Further mechanistic analyses demonstrated that IL-7 enhanced the expression of key migratory integrins (very late antigen-4 (VLA-4)) involved in T cell crossing of the blood-brain barrier endothelium. In addition, IL-7 upregulated the transcription of sphingosine-1-phosphate receptor 1 (S1P1), which has been shown to protect T cells from tumor-directed sequestration. Importantly for clinical translation, these findings were not just evaluated using lymphocytes from animals, but also using human T cells, including from both healthy donors and patients with glioblastoma. My dissertation work pertaining to IL-7’s role in VLA-4 modulation and T cell accumulation in intracranial glioma was accepted for publication in the Journal for Clinical Investigation with myself as first author.

To assess the treatment effect of novel combination therapies like BRiTE & ALT more definitively in the clinical setting, I also am a strong proponent for the expansion of tissue-based research studies. Here, we can evaluate early in the development process whether (1) therapy reaches tumor and (2) if therapy exerts its intended effect. During my doctoral research, I additionally became deeply involved in clinical research, writing articles as first author on rational combinatorial clinical trial design (published in Clinical Cancer Research) and the need to expand the use of tissue-based trials to correct the drug development paradigm in glioblastoma (published in Nature Medicine). 

Putting these concepts into practice, I wrote the trial protocol and was named as a sub-investigator/scientific lead for multiple clinical trials at Duke University. I initiated the development and execution of a tissue-based trial evaluating the pharmacokinetics (PK) and pharmacodynamics (PD) of evolocumab, a PCSK9 inhibitor in glioma (PEskE, NCT04937413, FDA IND: 156590). This trial was completed in 2024, finding that evolocumab has poor BBB uptake, but that tumor tissue with higher titers of evolocumab demonstrated expected PD changes (increased MHC-I expression and enhanced CD8+ T cell infiltration). I presented the initial findings of PEskE at the American Association of Neurosurgeons in 2024, where I received the Journal of Neuro-Oncology award and subsequently published our completed findings in Scientific Reports. Work is ongoing to explore combining evolocumab with BBB-opening therapies such as focused ultrasound. Building on this experience, I also designed and am currently working on a further tissue-based trial exploring the ability of fluoxetine to induce lysosomal stress in glioblastoma (FLiRT, NCT0563470, FDA IND: 163433). This study was based on pre-clinical research carried out at Stanford University in the laboratory of Dr. Paul Mischel, and which is now acting as a partner site for recruitment, alongside NYU Langone (under Dr. Erik Sulman) and UC San Diego (under Dr. David Piccioni).

From the beginning of my Ph.D., I was fortunate to receive mentorship from a large network within the brain tumor center. This included Dr. Peter E. Fecci, in whose lab the sequestration phenomenon was first described, and Dr. Mustafa Khasraw, who is the PI on numerous clinical trials, including the proposed first-in-human studies of BRiTE. After Dr. Sampson left Duke to take up a new role as Dean of Medicine at the University of Colorado, I joined Dr. Fecci’s laboratory with Dr. Khasraw as a co-advisor. Dr. Sampson remained on my committee throughout to provide ongoing mentorship and support. With the support and guidance of these outstanding physician-scientists, I completed my Ph.D. in 4 years, graduating in May 2025. 

These efforts and experiences are partially reflected in the following publications:

1. Singh K, Hotchkiss KM, Cook SL, Noldner P, Zhou Y, Moelker EM, Railton Co, Blandford EE, Puviindran BJ, Wallace SE, Norberg PK, Archer GE, Shaz BH, Ayasoufi K, Sampson JH, Khasraw M, Fecci PE. IL-7-mediated expansion of autologous lymphocytes increases CD8+ T Cell VLA-4 expression and accumulation within glioblastoma. J Clin Invest. 2025;135(12):e181471. PubMed Central PMCID: PMC12165802. 

2. Singh K, Batich KA, Wen PY, Tan AC, Bagley SJ, Lim M, Platten M, Colman H, Ashley DM, Chang SM, Rahman R, Galanis E, Mansouri A, Puduvalli VK, Reardon DA, Sahebjam S, Sampson JH, Simes J, Berry DA, Zadeh G, Cloughesy TF, Mehta MP, Piantadosi S, Weller M, Heimberger AB, Khasraw M. Designing Clinical Trials for Combination Immunotherapy: A Framework for Glioblastoma. Clin Cancer Res. 2022 Feb 15;28(4):585-593. PubMed Central PMCID: PMC9306329. Cited in SNO & EANO consensus guidelines on management of Glioblastoma in adults (Neuro-Oncology, 2025). 

3. Singh K, Hotchkiss KM, Parney IF, De Groot J, Sahebjam S, Sanai N, Platten M, Galanis E, Lim M, Wen PY, Minniti G, Colman H, Cloughesy TF, Mehta MP, Geurts M, Arrillaga-Romany I, Desjardins A, Tanner K, Short S, Arons D, Duke E, Wick W, Bagley SJ, Ashley DM, Kumthekar P, Verhaak R, Chalmers AJ, Patel AP, Watts C, Fecci PE, Batchelor TT, Weller M, Vogelbaum MA, Preusser M, Berger MS, Khasraw M. Correcting the drug development paradigm for glioblastoma requires serial tissue sampling. Nat Med. 2023 Oct;29(10):2402-2405. PubMed PMID: 37488293. Cited in SNO & EANO consensus guidelines on management of Glioblastoma in adults (Neuro-Oncology, 2025). 

4. Singh K, Foster MW, Violette MJ, Corcoran AM, Hotchkiss KM, Railton CO, Blandford EE, Blethen KE, Thomas EL, McIntosh WC, Ashley DM, Desjardins A, Friedman HS, Johnson MO, Friedman A, Keir S, Buckley ED, Herndon JE, McLendon RE, Sampson JH, Calabrese E, Lopez GY, Grant GA, Patel AP, Gregory SG, Li CY, Fecci PE, Khasraw M. A surgical window of opportunity trial evaluating the effect of the PCSK9 inhibitor evolocumab on tumoral MHC-I expression and CD8+ infiltration in glioma. Sci Rep. 2025. DOI: 10.1038/s41598-025-21064-9

Ongoing Research Support

NCT05634707: FLIRT: Evaluation of Fluoxetine and Cytotoxic Lysosomal Stress in Glioma

Role: Protocol design, pre-clinical and correlative science lead

NCT04937413: PESKE: The PCSK9i Inhibitor Evolocumab - a Surgical Trial of Pharmacodynamics and Kinetics Evaluation

Role: Sub-Investigator of ongoing clinical trial & protocol design, first-author on ultimate publication

NCT04903795: BRiTE: Bispecific T Cell Engager BRiTE for Patients with Grade IV Malignant Glioma

Role: Protocol design, pre-clinical, manufacturing and correlative science lead

Positions and Scientific Appointments

05/2025-current, Adaptin Bio & Duke University: Chief Scientific Consultant leading EGFRvIII BRiTE translation

08/2021-05/2025, Duke University: Biomedical Engineering PhD 

10/2020-current, University of Dundee: Honorary Clinical Lecturer

10/2020-07/2021, Duke University Medical Center: Research Scholar

08/2018-09/2020, National Health Service, Cardiff, UK: Surgical Resident

08/2016-08/2020, National Health Service, Dundee, UK: Academic Intern (medicine and surgery)

Honors

2024    Journal of Neuro-Oncology Award, American Association of Neurological Surgery ASM

2023    Preparing Future Faculty Fellow, Duke University

2023    BME Outstanding Teaching Assistant Award, Duke University

2023    Paul and Lauren Ghaffari Graduate Oncology Research Fellowship, Duke University

2020    Postdoctoral and Early Career Researcher Exchange award, Scottish Imaging Network

2018    Best oral poster presentation in session, Association of Surgeons of Great Britain and Ireland

2017    Best poster prize in healthcare work undertaken by postgraduates, Royal College of Physicians of Edinburgh

Other Experience and Professional Memberships

General Medical Council, UK

Royal College of Surgeons, England, UK

Congress of Neurological Surgeons, USA

Society of Neuro-Oncology, USA

Society for Immunotherapy of Cancer, USA

Research Projects

My research experience includes both clinical and pre-clinical research, investigating how to best optimize current approaches undertaken in the hospital environment, while exploring novel pre-clinical strategies for treating malignant brain tumors

Evaluating novel mechanisms of drug delivery across the blood-brain barrier (BBB)

As a core component of my pre-clinical PhD research, I have been developing a novel mechanism for drug delivery into the CNS, by using stimulated T cells which can cross the BBB with macromolecules (such as antibody-based therapies) bound to their surface. I presented my initial findings at the annual meeting of the Society for Neuro-Oncology in 2021 and continue to refine the process by which T cells are stimulated, in order to best traverse the BBB