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| Cytopathic effects and local immune responses in repeated neoadjuvant HSV-tk + ganciclovir gene therapy for prostate cancer |
Nobuyuki Yanagisawaa,b,*( ),Takefumi Satohc,d,Ken-ichi Tabatac,Hideyasu Tsumurac,Yasutomo Nasue,Masami Watanabee,f,Timothy C. Thompsong,Isao Okayasub,h,Yoshiki Murakumob,Shiro Babac,Masatsugu Iwamurac
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a Department of Pathology, St. Marianna University School of Medicine Yokohama-City Seibu Hospital, Yokohama, Kanagawa, Japan
b Department of Pathology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
c Department of Urology, Kitasato University School of Medicine, Sagamihara, Kanagawa, Japan
d Takefumi Satoh Prostate Clinic, Machida, Tokyo, Japan
e Department of Urology, Okayama University, Okayama, Japan
f Center for Innovative Clinical Medicine, Okayama University Hospital, Okayama, Japan
g Department of Genitourinary Medical Oncology - Research, Division of Cancer Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA
h Division of Nutrition, School of Health Care, Kiryu University, Midori-City Gunma, Japan |
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Abstract Objective: Cytopathic effects and local immune response were analyzed histologically in prostatic cancer (PCa) with in situ herpes simplex virus-thymidine kinase (HSV-tk)/ganciclovir (GCV) gene therapy (GT).
Methods: Four high-risk PCa patients who received HSV-tk/GCV GT were investigated. After two cycles of intraprostatic injection of HSV-tk and administration of GCV, radical prostatectomy was performed. Formalin-fixed, paraffin-embedded sections were evaluated using immunohistochemistry. PCa with hormone therapy (HT, n=3) or without neoadjuvant therapy (NT, n=4) that were equivalent in terms of risk were also examined as reference. Immunoreactively-positive cells were counted in at least three areas in cancer tissue. Labeling indices (LI) were calculated as percentage values.
Results: ssDNA LI in GT increased, indicating apoptosis, as well as tumor-infiltrating lymphocytes and CD68-positive macrophages, compared with their biopsies. GT cases showed significantly higher numbers of single-stranded DNA (ssDNA) LI, CD4/CD8-positive T cells and CD68-positive macrophages including M1/M2 macrophages than HT or NT cases. However, there was no significant difference in CD20-positive B cells among the types of case. There were strong correlations between CD8+ T cells and CD68+ macrophages (ρ=0.656, p<0.0001) as well as CD4+ T cells and CD20+ B cells (ρ=0.644, p<0.0001) in PCa with GT.
Conclusions: Enhanced cytopathic effect and local immune response might be indicated in PCa patients with HSV-tk/GCV gene therapy.
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Received: 10 April 2019
Available online: 16 June 2020
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Corresponding Authors:
Nobuyuki Yanagisawa
E-mail: czp05616@nifty.com
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Serum PSA levels in prostatic cancer patients treated with HSV-tk/GCV gene therapy. The PSA levels decreased after increasing temporarily at HSV-tk injections. PSA, prostate-specific antigen; GCV, ganciclovir; HSV-tk, herpes-simplex virus-tyrosine kinase.
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| Patient No. | Entry age | Baseline PSA, ng/mL | Biopsy Gleason Score | Positive Cores | cStage | Kattan Score | Therapy | pStage | Tumor affected, % | PSA reduction rate, % | Fever | Liver dysfunction (AST) | Liver dysfunction (ALT) | Thrombo-cytopenia | Prolonged APTT | Detection of ADV (Day 0, 2, 7, 16, 21, 28, 42, 56) | Maximum anti-ADV antibody titer (days) | | Blood | Urine | Nasal cavity | | 1 | 70 | 13.6 | 3+3=6 | 6/8 | T2c | 117 | RP | T2c | 15.6 | 24.1 | Grade 2 | Grade 1 | Grade 1 | | | - | - | - | x512 (28) | | 2 | 63 | 5.89 | 3+5=8 | 6/22 | T2b | 116 | RP | T2a | 9.5 | 30.0 | Grade 2 | Grade 3 | Grade 3 | Grade 1 | | - | - | - | x256 (28) | | 3 | 60 | 42.1 | 4+3=7 | 3/6 | T2c | 166 | HT | - | - | 32.0 | Grade 1 | Grade 1 | | | Grade 1 | - | - | - | x16 (42) | | 4 | 68 | 7.43 | 4+3=7 | 6/10 | T2b | 123 | RP | T2b | 52.7 | 27.7 | Grade 1 | Grade 1 | Grade 1 | Grade 1 | | - | - | - | x32 (14) | | 5 | 71 | 17.76 | 3+3=6 | 2/8 | T2b | 125 | RP | T3a | 22.8 | 1.7 | Grade 2 | Grade 1 | | | | - | - | - | x1024 (14) |
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Clinicopathologic characteristics of prostatic cancer patients treated with HSV-tk/GCV gene therapy.
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Histological results of prostatic cancer patients with HSV-tk gene therapy. (A) The cancer cells showed glandular dissolution, nuclear pyknosis and apoptotic bodies (arrows). Inflammatory cell infiltration was also observed. (B) The cancer cells almost lost PSA positivity. (C) Cytokeratin (CAM5.2) was still focally positive. (D) ssDNA indicating apoptosis was scattered positive in cancer cells. (E-I) Increased CD8+ T cells (E), CD68+ (F) and CD11c+ M1 macrophages (H) were observed in tumor, while CD20+ B cells (G) and CD163+ M2 macrophages (I) were a few. Original magnification, ×400. Scale bars, 50 μm. PSA, prostate-specific antigen; HSV-tk, herpes-simplex virus-tyrosine kinase.
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Comparison of cytopathic effects and local immune responses between prostatic cancer and non-cancerous lesions in patients with HSV-tk/GCV gene therapy. The cancer cells (T) increased ssDNA labelling index compared with non-cancerous lesions (N). In contrast, the non-cancerous legions (N) tended to show more inflammatory cell infiltration than intratumoral legions (T). Comparisons between groups were conducted with the Manne-Whitney U test. p-Value <0.05 was considered significant. HSV-tk, herpes-simplex virus-tyrosine kinase; ssDNA, single-stranded DNA.
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Comparison of cytopathic effects and local immune responses between biopsy and radical prostatectomy specimens in prostatic cancer patients with HSV-tk/GCV gene therapy. ssDNA labelling index and intratumoral CD8+ T cells, CD68+ macrophages and CD20+ B cells in the radical prostatectomy specimens (RP) treated with gene therapy were significantly increased compared with their biopsies (BP) before the treatment. p-Value <0.05 was considered significant. HSV-tk, herpes-simplex virus-tyrosine kinase.
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Cytopathic effects and local immune responses in prostatic cancer with or without neoadjuvant treatment. (A) CD4, CD8, CD68-positive cells significantly increased in the prostatic cancers of patients with HSV-tk/GCV gene therapy (GT) compared with hormone therapy (HT) or no neoadjuvant therapy (NT) patients. (B) ssDNA labelling index (LI) also increased, while Ki-67 LI was not significant in GT patients. Both CD11c+ (M1 macrophages) and CD163+ (M2 macrophages) cells also increased, and M1 macrophages were predominant in GT tumors. Statistical analysis was done using Fisher protected least significant difference test. p-Value<0.05 was considered significant.
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| marker | ρa | p-Valueb | | ssDNA/CD4 | -0.322 | 0.0271 | | ssDNA/CD8 | -0.084 | 0.5642 | | ssDNA/CD68 | -0.193 | 0.1853 | | ssDNA/CD20 | -0.474 | 0.0012 | | CD4/CD8 | 0.466 | 0.0014 | | CD4/CD68 | 0.483 | 0.0009 | | CD4/CD20 | 0.644 | <0.0001 | | CD8/CD68 | 0.656 | <0.0001 | | CD8/CD20 | 0.381 | 0.0091 | | CD68/CD20 | 0.392 | 0.0072 |
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Correlation coefficients for ssDNA and intratumoral inflammatory cells in prostatic cancer patients with HSK-tk/GCV gene therapy.
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