In Vivo 19F-Magnetic Resonance Imaging of Adoptively Transferred NK Cells

Somanchi, Srinivas S.; Kennis, Bridget; Gopalakrishnan, Vidya; Lee, Dean A.; Bankson, James A.

doi:10.1007/978-1-4939-3684-7_27

Cited by 19 publications

(20 citation statements)

References 28 publications

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“…Cell labeling in culture is generally performed by simple co-incubation with PFC as another factor in the media, followed by a wash step. Labeling periods range from several hours [ 21 , 34 – 36 ] to a day or more [ 37 – 39 ] to allow for endocytic uptake to occur. Determinants of obtainable PFC cell uptake include (i) dose of PFC in media, (ii) cell cytoplasmic volume and (iii) phagocytic properties of cells.…”

Section: Introductionmentioning

confidence: 99%

“…Similarly, NK cells labeled with PFPE nanoemulsion exhibited unaltered viability and phenotype [ 37 ]. Somanchi et al published a detailed protocol for expansion and PFPE labeling of NK cells [ 36 ]. Cytotoxicity of labeled NK cells against cancer cells in vitro was comparable to non-labeled cells, and cytokine and perforin secretion was preserved [ 36 , 37 ] (Table 1 ).…”

Section: Introductionmentioning

confidence: 99%

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Fluorine-19 MRI for detection and quantification of immune cell therapy for cancer

Chapelin¹,

Capitini²,

Ahrens³

2018

j. immunotherapy cancer

104

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Over the past two decades, immune cell therapy has emerged as a potent treatment for multiple cancers, first through groundbreaking leukemia therapy, and more recently, by tackling solid tumors. Developing successful therapeutic strategies using live cells could benefit from the ability to rapidly determine their in vivo biodistribution and persistence. Assaying cell biodistribution is unconventional compared to traditional small molecule drug pharmacokinetic readouts used in the pharmaceutical pipeline, yet this information is critical towards understanding putative therapeutic outcomes and modes of action. Towards this goal, efforts are underway to visualize and quantify immune cell therapy in vivo using advanced magnetic resonance imaging (MRI) techniques. Cell labeling probes based on perfluorocarbon nanoemulsions, paired with fluorine-19 MRI detection, enables background-free quantification of cell localization and survival. Here, we highlight recent preclinical and clinical uses of perfluorocarbon probes and 19F MRI for adoptive cell transfer (ACT) studies employing experimental T lymphocytes, NK, PBMC, and dendritic cell therapies. We assess the forward looking potential of this emerging imaging technology to aid discovery and preclinical phases, as well as clinical trials. The limitations and barriers towards widespread adoption of this technology, as well as alternative imaging strategies, are discussed.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fluorine-19 MRI for detection and quantification of immune cell therapy for cancer

Chapelin¹,

Capitini²,

Ahrens³

2018

j. immunotherapy cancer

104

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“…Fluorine-19 MRI methods have shown promise for the detection of cell therapy products post-transfer, 12,[14][15][16]18,19,38 inflammatory infiltrates, [39][40][41][42][43] and molecular targets 44 in vivo in preclinical models. Moreover, first-generation 19 F probes based on PFC nanoemulsions have been used in a pilot clinical trial.…”

Section: Discussionmentioning

confidence: 99%

“…12,13 Fluorine-19-based MRI nanoemulsion probes are an option for non-invasively imaging of cell populations. [14][15][16][17][18][19][20] The 19 F nuclei have high intrinsic sensitivity, with 89% relative sensitivity compared to 1 H. De minimis endogenous 19 F in the body ensures that any MRI signals collected are from the introduced tracer probe. F-dense perfluorocarbon (PFC) molecules are often used to form nanoemulsion imaging probes that can be endocytosed by cells.…”

Section: Introductionmentioning

confidence: 99%

Cell penetrating peptide functionalized perfluorocarbon nanoemulsions for targeted cell labeling and enhanced fluorine‐19 MRI detection

Hingorani

Chapelin

Stares

et al. 2019

Magnetic Resonance in Med

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Purpose: A bottleneck in developing cell therapies for cancer is assaying cell biodistribution, persistence, and survival in vivo. Ex vivo cell labeling using perfluorocarbon (PFC) nanoemulsions, paired with 19 F MRI detection, is a noninvasive approach for cell product detection in vivo. Lymphocytes are small and weakly phagocytic limiting PFC labeling levels and MRI sensitivity. To boost labeling, we designed PFC nanoemulsion imaging probes displaying a cell-penetrating peptide, namely the transactivating transcription sequence (TAT) of the human immunodeficiency virus. We report optimized synthesis schemes for preparing TAT co-surfactant to complement the common surfactants used in PFC nanoemulsion preparations. Methods: We performed ex vivo labeling of primary human chimeric antigen receptor (CAR) T cells with nanoemulsion. Intracellular labeling was validated using electron microscopy and confocal imaging. To detect signal enhancement in vivo, labeled CAR T cells were intra-tumorally injected into mice bearing flank glioma tumors. Results: By incorporating TAT into the nanoemulsion, a labeling efficiency of ~10 12 fluorine atoms per CAR T cell was achieved that is a >8-fold increase compared to nanoemulsion without TAT while retaining high cell viability (~84%). Flow cytometry phenotypic assays show that CAR T cells are unaltered after labeling with TAT nanoemulsion, and in vitro tumor cell killing assays display intact cytotoxic function.The 19 F MRI signal detected from TAT-labeled CAR T cells was 8 times higher than cells labeled with PFC without TAT. Conclusion: The peptide-PFC nanoemulsion synthesis scheme presented can significantly enhance cell labeling and imaging sensitivity and is generalizable for other targeted imaging probes.

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“…Epigenetic drugs of various targets have shown promising anti-tumor effects in pre-clinical DIPG models, but these agents have not yet been combined with clinically relevant immunotherapy regimens. NK cell administration is effective against malignant gliomas 27 , can be grown without autologous sources of leukocytes 28 , and has already begun clinical trials using our exact NK-expansion methodology 29 . Furthermore, combination therapy with LSD1 inhibitors is desirable due to the demonstrated selectivity of LSD1i-induced DIPG cell death versus normal astrocytes.…”

Section: Discussionmentioning

confidence: 99%

Pharmacologic inhibition of lysine specific demethylase-1 (LSD1) as a therapeutic and immune-sensitization strategy in diffuse intrinsic pontine glioma (DIPG)

Bailey

Romero²,

Becher³

et al. 2019

Preprint

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BackgroundDiffuse intrinsic pontine glioma (DIPG) is an incurable pediatric brain tumor. Mutations in the H3 histone tail (H3.1/3.3-K27M) are a feature of DIPG, potentially rendering them therapeutically sensitive to small-molecule inhibition of chromatin modifiers. Pharmacological inhibition of lysine specific demethylase-1 (LSD1) shows promise in pediatric cancers such as Ewing’s sarcoma, but has not been investigated in DIPG, which was the aim of our study.MethodsPatient-derived DIPG cell lines and pediatric high-grade glioma (pHGG) datasets were used to evaluate effects of several LSD1 inhibitors on selective cytotoxicity and immune gene expression. Immune cell cytotoxicity was assessed in DIPG cells treated with LSD1 inhibitors and informatics platforms were used to determine immune infiltration of pHGG and impact on survival.ResultsSelective cytotoxicity and an immunogenic gene signature was established in DIPG lines using several clinically-relevant LSD1 inhibitors. Pediatric high-grade glioma patient sequencing data demonstrated survival benefit using this LSD1-dependent gene signature. On-target binding of catalytic LSD1 inhibitors was confirmed in DIPG and pre-treatment of DIPG with these inhibitors increased lysis by natural killer (NK) cells. CIBERSORT analysis of patient data confirmed NK infiltration is beneficial to patient survival while CD8 T-cells are negatively prognostic. Catalytic LSD1 inhibitors are non-perturbing to NK cells while scaffolding LSD1 inhibitors are toxic to NK cells and do not induce the gene signature in DIPG cells.ConclusionsLSD1 inhibition using catalytic inhibitors are both selectively cytotoxic and promote an immune gene signature that is associated with NK cell killing, representing a therapeutic opportunity for pHGG.Key pointsLSD1 inhibition using several clinically relevant compounds is selectively cytotoxic in DIPG.An LSD1-controlled gene signature predicts survival in pediatric high-grade glioma patients.LSD1 inhibition enhances NK cell cytotoxicity against DIPG with correlative genetic biomarkers.Importance of the studyThis is the first study to evaluate inhibition of LSD1 in a uniformly lethal type of pediatric brain tumor: DIPG. We demonstrate selective cytotoxicity of several clinically relevant compounds against patient derived DIPG cells, and identify an immune gene signature that is upregulated in DIPG cells by catalytic inhibitors of LSD1. This immune gene signature is predictive of prognosis in pHGG, consistent with the rationale of promoting this signature through LSD1 inhibition. NK cell killing of DIPG is enhanced by LSD1 inhibition, providing functional confirmation of this gene signature, and represents the first report of LSD1 inhibition promoting NK cell cytotoxicity of cancer cells. Given the poor prognosis of pHGGs and lack of effective treatments, our results suggest use of LSD1 inhibition as a single agent or in combination with NK cell therapy may be a safe and efficacious strategy.

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In Vivo 19F-Magnetic Resonance Imaging of Adoptively Transferred NK Cells

Cited by 19 publications

References 28 publications

Fluorine-19 MRI for detection and quantification of immune cell therapy for cancer

Fluorine-19 MRI for detection and quantification of immune cell therapy for cancer

Cell penetrating peptide functionalized perfluorocarbon nanoemulsions for targeted cell labeling and enhanced fluorine‐19 MRI detection

Pharmacologic inhibition of lysine specific demethylase-1 (LSD1) as a therapeutic and immune-sensitization strategy in diffuse intrinsic pontine glioma (DIPG)

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