Monitoring Drug-Induced γH2AX as a Pharmacodynamic Biomarker in Individual Circulating Tumor Cells

Wang, Lihua H.; Pfister, Thomas D.; Parchment, Ralph E.; Kummar, Shivaani; Rubinstein, Larry; Gutierrez, Martin; Murgo, Anthony J.; Tomaszewski, Joseph E.; Doroshow, James H.; Kinders, Robert J.

doi:10.1158/1078-0432.ccr-09-2799

Cited by 89 publications

(83 citation statements)

References 38 publications

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“…Cells originating from tumors receiving site-directed radiation are marked by ionizing radiation-induced DNA damage, including tumor and stromal cells (18)(19)(20)(21)(22)(23)(24). Thus, we expect that circulating cells that originate at the tumor site during radiotherapy should have evidence of DNA damage, such as ionizing radiation-induced foci (IRIF), which can be visualized with RAD50 (18)(19)(20)(21)(22)(23)(24).…”

Section: Introductionmentioning

confidence: 99%

Sequential Tracking of PD-L1 Expression and RAD50 Induction in Circulating Tumor and Stromal Cells of Lung Cancer Patients Undergoing Radiotherapy

Adams

et al. 2017

Clinical Cancer Research

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Purpose: Evidence suggests that PD-L1 can be induced with radiotherapy and may be an immune escape mechanism in cancer. Monitoring this response is limited, as repetitive biopsies during therapy are impractical, dangerous, and miss tumor stromal cells. Monitoring PD-L1 expression in both circulating tumor cells (CTCs) and circulating stromal cells (CStCs) in blood-based biopsies might be a practical alternative for sequential, noninvasive assessment of changes in tumor and stromal cells.Experimental Design: Peripheral blood was collected before and after radiotherapy from 41 patients with lung cancer, as were primary biopsies. We evaluated the expression of PD-L1 and formation of RAD50 foci in CTCs and a CStC subtype, cancer-associated macrophage-like cells (CAMLs), in response to DNA damage caused by radiotherapy at the tumor site.Results: Only 24% of primary biopsies had sufficient tissue for PD-L1 testing, tested with IHC clones 22c3 and 28-8. A CTC or CAML was detectable in 93% and 100% of samples, prior to and after radiotherapy, respectively. RAD50 foci significantly increased in CTCs (>7Â, P < 0.001) and CAMLs (>10Â, P ¼ 0.001) after radiotherapy, confirming their origin from the radiated site. PD-L1 expression increased overall, 1.6Â in CTCs (P ¼ 0.021) and 1.8Â in CAMLs (P ¼ 0.004): however, individual patient PD-L1 expression varied, consistently low/negative (51%), consistently high (17%), or induced (31%).Conclusions: These data suggest that RAD50 foci formation in CTCs and CAMLs may be used to track cells subjected to radiation occurring at primary tumors, and following PD-L1 expression in circulating cells may be used as a surrogate for tracking adaptive changes in immunotherapeutic targets.

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Section: Introductionmentioning

confidence: 99%

Sequential Tracking of PD-L1 Expression and RAD50 Induction in Circulating Tumor and Stromal Cells of Lung Cancer Patients Undergoing Radiotherapy

Adams

et al. 2017

Clinical Cancer Research

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“…There are reports of cytological γH2AX assay, using circulating tumour cells (CTCs) 11, 12, 13. In one study, a threshold in the γH2AX FACS signal was used to categorise cells as positive or negative.…”

Section: Discussionmentioning

confidence: 99%

“…In one study, a threshold in the γH2AX FACS signal was used to categorise cells as positive or negative. The proportion of positive cells increased from 0–7% to 22–64% with chemotherapy treatment, and a mean of only 38 cells were examined 11. The extent to which FNA or CTC represents the tumour as a whole is uncertain, although FNA can potentially sample across a whole tumour mass, rather than merely cells that are able to enter the circulation.…”

Section: Discussionmentioning

confidence: 99%

γH2AX expression in cytological specimens as a biomarker of response to radiotherapy in solid malignancies

Shah

Boghozian

Kartsonaki

et al. 2015

Diagnostic Cytopathology

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Many anticancer treatments, including radiotherapy, act by damaging DNA and hindering cell function and proliferation. H2AX is a histone protein directly associated with DNA that is phosphorylated to produce γH2AX that accumulates in foci in an early response to DNA double‐strand breaks, the most deleterious lesion caused by anticancer therapy. This study reports a γH2AX detection assay that has the potential to be used as a biomarker of response to guide cancer treatment. γH2AX immunostaining was applied to tumour cell specimens obtained using fine needle aspiration (FNA). Liquid‐based cytology and direct smear cytology methods were evaluated and immunostaining protocols established using FNA samples from five cancer patients. The assay was then applied to three patients before and after radiotherapy. Results demonstrate induction of γH2AX foci following treatment, persisting for as long as one week after therapy. Immunostaining for γH2AX has been successfully applied to FNA samples, providing an opportunity to evaluate γH2AX as a treatment response marker in cancer. Diagn. Cytopathol. 2016;44:141–146. © 2015 The Authors Diagnostic Cytopathology Published by Wiley Periodicals, Inc.

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“…8 Note that other diagnostic markers could be readily incorporated into the HMSS platform, including DNA damage markers (e.g., histone cH2AX) and signaling proteins (e.g., phosphorylated EGFR). 36,37 Such multiplexed assays on CTC molecular status would be of significant clinical importance in monitoring and rationalizing patient treatment.…”

Section: Molecular Profiling On Individual Cancer Cellsmentioning

confidence: 99%

Rare cell isolation and profiling on a hybrid magnetic/size-sorting chip

et al. 2013

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We present a hybrid magnetic/size-sorting (HMSS) chip for isolation and molecular analyses of circulating tumor cells (CTCs). The chip employs both negative and positive cell selection in order to provide high throughput, unbiased CTC enrichment. Specifically, the system utilizes a self-assembled magnet to generate high magnetic forces and a weir-style structure for cell sorting. The resulting device thus can perform multiple functions, including magnetic depletion, size-selective cell capture, and on-chip molecular staining. With such capacities, the HMSS device allowed one-step CTC isolation and single cell detection from whole blood, tested with spiked cancer cells. The system further facilitated the study of individual CTCs for heterogeneity in molecular marker expression.

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Monitoring Drug-Induced γH2AX as a Pharmacodynamic Biomarker in Individual Circulating Tumor Cells

Cited by 89 publications

References 38 publications

Sequential Tracking of PD-L1 Expression and RAD50 Induction in Circulating Tumor and Stromal Cells of Lung Cancer Patients Undergoing Radiotherapy

Sequential Tracking of PD-L1 Expression and RAD50 Induction in Circulating Tumor and Stromal Cells of Lung Cancer Patients Undergoing Radiotherapy

γH2AX expression in cytological specimens as a biomarker of response to radiotherapy in solid malignancies

Rare cell isolation and profiling on a hybrid magnetic/size-sorting chip

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