Donor-Derived Keratinocytes in Actinic Keratosis and Squamous Cell Carcinoma in Patients with Kidney Transplant
2012; Elsevier BV; Volume: 133; Issue: 4 Linguagem: Inglês
10.1038/jid.2012.422
ISSN1523-1747
AutoresL. Verneuil, Mariana Varna, Christophe Lebœuf, Louis François Plassa, Morad Elbouchtaoui, Irmine Ferreira, Fatiha Bouhidel, Marie‐Noelle Péraldi, Célèste Lebbé, Philippe Ratajczak, Anne Janin,
Tópico(s)Corneal Surgery and Treatments
Resumoactinic keratosis squamous cell carcinoma TO THE EDITOR The homing ability of hematopoietic stem cells has enabled their therapeutic use. We have previously shown in allogeneic hematopoietic stem-cell recipients that donor-derived cells can be detected in skin in acute graft-versus-host disease (Murata et al., 2007Murata H. Janin A. Leboeuf C. et al.Donor-derived cells and human graft-versus-host disease of the skin.Blood. 2007; 109: 2663-2665Crossref PubMed Scopus (52) Google Scholar) and in squamous cell carcinoma (SCC) of the oral mucosa (Janin et al., 2009Janin A. Murata H. Leboeuf C. et al.Donor-derived oral squamous cell carcinoma after allogeneic bone marrow transplantation.Blood. 2009; 113: 1834-1840Crossref PubMed Scopus (53) Google Scholar). In kidney transplant recipients, population-based studies show an increased risk of skin SCC (Moloney et al., 2006Moloney F.J. Comber H. O'Lorcain P. et al.A population-based study of skin cancer incidence and prevalence in renal transplant recipients.Br J Dermatol. 2006; 154: 498-504Crossref PubMed Scopus (367) Google Scholar; Wisgerhof et al., 2010Wisgerhof H.C. Edelbroek J.R. de Fijter J.W. et al.Subsequent squamous- and basal-cell carcinomas in kidney-transplant recipients after the first skin cancer: cumulative incidence and risk factors.Transplantation. 2010; 89: 1231-1238Crossref PubMed Scopus (84) Google Scholar), which are remarkable for: (i) their incidence, 250 times higher than in the normal population (Mackenzie et al., 2010Mackenzie K.A. Wells J.E. Lynn K.L. et al.First and subsequent nonmelanoma skin cancers: incidence and predictors in a population of New Zealand renal transplant recipients.Nephrol Dial Transplant. 2010; 25: 300-306Crossref PubMed Scopus (49) Google Scholar), (ii) their prevalence over basal cell carcinoma, (iii) the fact that they are multiple, and (iv) their association with multiple actinic keratosis (AK). Donor-derived cells have been identified in one case of basal cell carcinoma (Aractingi et al., 2005Aractingi S. Kanitakis J. Euvrard S. et al.Skin carcinoma arising from donor cells in a kidney transplant recipient.Cancer Res. 2005; 65: 1755-1760Crossref PubMed Scopus (76) Google Scholar). Here we address the question of the presence of donor-derived cells in skin SCC and in associated AK in kidney transplant recipients. In three patients with AK and SCC who had received sex-mismatched kidney transplants, and for whom remaining tissue samples and DNA from donor and recipient were available, we conducted a chimerism study using four independent methods, FISH for X and Y chromosome detection, ZFYqPCR, polymorphic microsatellite analyses, and HRM on mitochondrial DNA (Supplementary Data online). The study was approved by the Institutional Review Board of Hôpital Saint-Louis (Paris, France) and written informed consent was obtained according to the Helsinki Declaration. Download .pdf (.03 MB) Help with pdf files Supplementary Data AK and SCC lesions included in this study were not contiguous, but 5 to 10cm apart. XY-FISH combined with cytokeratin on the same tissue section showed cytokeratin-positive cells with the donor genotype in two female recipients of male kidney transplants. The percentage of chimeric cells was 4% in the SCC and 2% in the two AKs studied for Patient 1 (Figure 1a), and 5% in the SCC and 3 and 0% in the two AKs studied for Patient 2 (Table 1). Chimeric XY cells were distributed in the basal layer in AK and in the basal layer and the invasive areas in SCC. No chimeric cell was found in normal surrounding skin. The third patient with sex-mismatched kidney transplant (Patient 3) was male, and no chimeric XX cytokeratin-positive cell was found in his SCC or AK.Table 1XY-FISH data in AK and SCC of three sex-mismatched kidney transplant recipientsSexKidney transplantationSkin tumorXY FISHRecipient/donorAge at transplant (years)Timelapse transplant– tumor (years)SiteTypeKeratinocytes from basal and suprabasal layersXXXYTotalXX cells, %XY cells, %% Chimeric cells, corrected1Tumor samples from two patients with SCC and AK without kidney tranplantation setting (control) were examined. To determine the efficiency of sex chromosome detection in basal and suprabasal layers of SCC and AK, a FISH-XY protocol was applied. Tumor sections from the two patients with SCC and AK were analyzed and the percent detection averaged for either SCC {((79:127)+(72:112)x100):2=63.25%} or AK {((102:147)+(101:141)x100):2=70.5%}. The normalization factor was derived by dividing 100% by the average percent for either SCC (100:63.25=1.58) or AK (100:70.5=1.42). Here the normalization factor was 1.42 and 1.58 for XX and XY cells detections of AK and SCC, respectively.Patients with skin tumor after sex-mismatched kidney transplantXY cells 1F/M4314ForeheadSCC66410761.73.75.912ForeheadAK82212167.81.72.312CheekAK68211758.11.72.4 2F/M463NeckSCC6059861.25.18.13NeckAK88313266.72.33.31NeckAK95014167.40.00XX cells 3M/F595NoseSCC0711190.059.704NoseAK061940.064.90Patient with skin tumor after sex-matched kidney transplantationM/M437ForeheadSCC052910.057.17ForeheadAK0661090.060.6Patients with skin tumor without kidney transplantation (control)MNoseSCC0791270.062.2CheekAK01021470.069.4MLegSCC0721120.064.3LegAK01011410.071.61 Tumor samples from two patients with SCC and AK without kidney tranplantation setting (control) were examined. To determine the efficiency of sex chromosome detection in basal and suprabasal layers of SCC and AK, a FISH-XY protocol was applied. Tumor sections from the two patients with SCC and AK were analyzed and the percent detection averaged for either SCC {((79:127)+(72:112)x100):2=63.25%} or AK {((102:147)+(101:141)x100):2=70.5%}. The normalization factor was derived by dividing 100% by the average percent for either SCC (100:63.25=1.58) or AK (100:70.5=1.42). Here the normalization factor was 1.42 and 1.58 for XX and XY cells detections of AK and SCC, respectively. Open table in a new tab As donor-derived cells were only found in female recipients of male kidney transplants, we performed a qPCR for ZFY gene on AK and SCC tissue sections, which confirmed the presence of the Y chromosome in Patient 1 SCC and two AKs (Figure 1b) and Patient 2 SCC and one of the two AKs studied. To check these chromosomal results using completely different methods, which also associated molecular markers and morphological selection of cell populations, we performed microsatellite PCR and HRM analyses on laser-microdissected cells. Laser microdissection, performed on tissue sections after immunostaining of inflammatory cells, enabled us to collect only keratinocytes from the basal layer in AK and keratinocytes from the basal layer and from invasive areas in the dermis in SCC (Figure 1c). The polymorphic microsatellite marker analyses performed on laser-microdissected cells confirmed the presence of cells of donor origin in the SCC and two AKs of Patient 1, using four different dinucleotide repeats: D3S3611, D5S2095, D8S1820, and D9S162 (as shown for D3S3611 in Figure 1d). It also confirmed the presence of cells of donor origin in the SCC and one AK of Patient 2, using three different dinucleotide repeats: D3S1597, D5S2095, and D8S1820. Polymorphic microsatellite analyses D3S3611, D8S1820, and D17S1879 showed recipient origin for Patient 3 SCC and AK. We performed mitochondrial DNA HRM in the same samples of laser-microdissected keratinocytes from AK and SCC. This second independent method also showed the presence of cells of donor origin in the SCC and two AKs of Patient 1 (Figure 1e), and in the SCC and one AK of Patient 2. The cells of the SCC and AK of Patient 3 were all of recipient origin. Altogether, we provide conclusive evidence for the presence of donor-derived basal keratinocytes in AK, and of donor-derived basal and invasive keratinocytes in skin SCC in two kidney transplant recipients. These data imply the homing of stem/progenitor cells from the kidney transplants to the recipients' skin. For the homing of stem/progenitor cells from adult kidney to the skin, no experimental data are available. Multipotent progenitor cells from Bowman's capsule in adult human kidney have been isolated and characterized (Sagrinati et al., 2006Sagrinati C. Netti G.S. Mazzinghi B. et al.Isolation and characterization of multipotent progenitor cells from the Bowman's capsule of adult human kidneys.J Am Soc Nephrol. 2006; 17: 2443-2456Crossref PubMed Scopus (573) Google Scholar), but a transfer of mesenchymal stem cells with the transplant cannot be excluded. Using combined XY-FISH and immunostaining on whole tissue sections enabled us to observe that chimeric cells were only present in SCC and AK, and not in normal peripheral skin. In addition, the donor-derived cells we identified in SCC were located in the basal layer and in invasive areas, which correspond to the "outer proliferating layer" where tumor-initiating cells have recently been characterized in skin SCC (Patel et al., 2012Patel G.K. Yee C.L. Terunuma A. et al.Identification and characterization of tumor-initiating cells in human primary cutaneous squamous cell carcinoma.J Invest Dermatol. 2012; 132: 401-409Crossref PubMed Scopus (43) Google Scholar). We also identified donor-derived cells in the basal layer of AK, which is usually considered as a benign disease, although molecular studies have shown frequent loss of heterozygozity (Rehman et al., 1994Rehman I. Quinn A.G. Healy E. et al.High frequency of loss of heterozygosity in actinic keratoses, a usually benign disease.Lancet. 1994; 344: 788-789Abstract PubMed Scopus (64) Google Scholar) and gene expression patterns are in favor of a spectrum of disease progression from normal human skin to AK and SCC (Padilla et al., 2010Padilla R.S. Sebastian S. Jiang Z. et al.Gene expression patterns of normal human skin, actinic keratosis, and squamous cell carcinoma: a spectrum of disease progression.Arch Dermatol. 2010; 146: 288-293Crossref PubMed Scopus (102) Google Scholar). In our cases, the concentration of chimeric cells in AK and SCC could be linked to the tissue remodeling accompanying disease progression, as stem/progenitor cells are recruited to the sites of skin injuries in experimental conditions (Zong et al., 2008Zong Z.W. Cheng T.M. Su Y.P. et al.Recruitment of transplanted dermal multipotent stem cells to sites of injury in rats with combined radiation and wound injury by interaction of SDF-1 and CXCR4.Radiat Res. 2008; 170: 444-450Crossref PubMed Scopus (21) Google Scholar), and tissue repair shares common mechanisms with stem-cell renewal in carcinogenesis (Beachy et al., 2004Beachy P.A. Karhadkar S.S. Berman D.M. Tissue repair and stem cell renewal in carcinogenesis.Nature. 2004; 432: 324-331Crossref PubMed Scopus (1019) Google Scholar). In the kidney transplant recipients we studied, the precise role of chimeric stem cells in skin SCC oncogenesis remains to be established, but the detection of chimeric cells at the stage of AK is in favor of a multistep process in the disease progression. Grants were received from Région-Ile-de-France, Cancéropôle Ile-de-France, INCa, and ANR. LV is a Société-Française-Dermatologie grant recipient. A. Swaine assisted with language editing. Supplementary material is linked to the online version of the paper at http://www.nature.com/jid
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