Identification of Small-Molecule Inhibitors Targeting Different Signaling Pathways in Cancer-Associated Fibroblast Reprogramming Under Tumor-Stroma Interaction.

Cancer-associated fibroblasts (CAFs) interact reciprocally with tumor cells through various signaling pathways in many cancer types including cutaneous squamous cell carcinoma (cSCC). Among normal fibroblast (NF) subtypes, papillary fibroblasts (PFs) and reticular fibroblasts (RFs) respond distinctly to tumor cell signaling, eventuating the differentiation of RFs, rather than PFs, into CAFs. The regulation of subtype differentiation in fibroblasts remains poorly explored. In this study, we assessed the differences between PFs, RFs, and CAFs, and examined the effects of small-molecule inhibitors targeting the TGFß, PI3K/AKT/mTOR, and NOTCH pathways on the tumor-promoting property of CAFs and CAF reprogramming in 2D and 3D cultures. Blocking TGFß and PI3K strongly deactivated and concurrently induced a PF phenotype in RFs and CAFs. 3D co-culturing a cSCC cell line MET2 with RFs or CAFs led to enhanced tumor invasion, "RF-CAF" transition and cytokine production, which were further repressed by blocking TGFß and PI3K/mTOR pathways, but not NOTCH pathway. In conclusion, the study identified biomarkers for PFs, RFs and CAFs, and displayed different effects of blocking key signaling pathways in CAFs and tumor cell-CAF interplay. These findings prompted a "CAF to PF" therapeutic strategy, and provided perspectives of using included inhibitors in CAF-based cancer therapy.

Skin disorders indicating peripheral arterial occlusive disease and chronic venous insufficiency in organ transplant recipients.

BACKGROUND: Peripheral arterial occlusive disease (PAOD) and chronic venous insufficiency (CVI) in organ transplant recipients (OTR) can lead to harmful outcomes. We made an inventory of cutaneous manifestations of PAOD and CVI in OTR in relation with diabetes and other potential risk factors. METHODS: A prospective study in a single center was performed. OTR (n = 112) were included at the outpatient clinic to investigate clinical signs of PAOD and CVI. The most commonly associated risk factors were determined. RESULTS: PAOD had been diagnosed in 15.6% and CVI in 30.0% of the patients. Diabetes was the cause of organ failure in 9.8% of the patients. Type 1 diabetes had been diagnosed in 8.9% and type 2 diabetes in 21.4% (59.1% new-onset diabetes after transplantation). Type 1 diabetes showed an increased risk for PAOD and limb amputation with hazard ratios of 11.0 (95%CI 3.0-40.2) and 9.1 (95%CI 1.4-58.6). Type 2 diabetes showed no increased risk. CONCLUSIONS: Patients with a history of type 1 diabetes were at high risk for PAOD even years after a simultaneous pancreas kidney transplantation and they should remain under close observation for PAOD even though they are supposedly "cured" from their diabetes to prevent a harmful outcome.

Metastasis Risk of Cutaneous Squamous Cell Carcinoma in Organ Transplant Recipients and Immunocompetent Patients.

Organ transplant recipients (OTRs) have a high incidence of cutaneous squamous cell carcinoma (cSCC), and immunosuppression has been reported to be an important risk factor for metastasis. The aim of this study was to identify the metastasis risk over a 10-year period for 593 patients with cSCC, of whom 134 were OTR and 459 were immunocompetent. Metastasis incidence rate was 1,046 (95% confidence interval (95% CI) 524-2,096) per 100,000 person years in OTR and 656 (95% CI; 388-1,107) in immunocompetent patients, yielding an incidence rate ratio of 1.6 (95% CI 0.67-3.81). In OTRs head/neck location, older age at transplantation and older age at diagnosis of first cSCC were associated with metastatic risk, and 7 out of 8 metastasized tumours were smaller than 2 cm. In immunocompetent patients tumour size and tumour depth were associated with metastasis. In conclusion, we were not able to demonstrate an increased incidence rate of metastasis in OTRs compared with immunocompetent patients. However, OTRs and immunocompetent patients differed with regard to risk factors for metastasis.

Human papillomavirus and posttransplantation cutaneous squamous cell carcinoma: A multicenter, prospective cohort study.

Organ transplant recipients (OTRs) have a 100-fold increased risk of cutaneous squamous cell carcinoma (cSCC). We prospectively evaluated the association between ß genus human papillomaviruses (ßPV) and keratinocyte carcinoma in OTRs. Two OTR cohorts without cSCC were assembled: cohort 1 was transplanted in 2003-2006 (n = 274) and cohort 2 was transplanted in 1986-2002 (n = 352). Participants were followed until death or cessation of follow-up in 2016. ßPV infection was assessed in eyebrow hair by using polymerase chain reaction-based methods. ßPV IgG seroresponses were determined with multiplex serology. A competing risk model with delayed entry was used to estimate cumulative incidence of histologically proven cSCC and the effect of ßPV by using a multivariable Cox regression model. Results are reported as adjusted hazard ratios (HRs). OTRs with 5 or more different ßPV types in eyebrow hair had 1.7 times the risk of cSCC vs OTRs with 0 to 4 different types (HR 1.7, 95% confidence interval 1.1-2.6). A similar risk was seen with high ßPV loads (HR 1.8, 95% confidence interval 1.2-2.8). No significant associations were seen between serum antibodies and cSCC or between ßPV and basal cell carcinoma. The diversity and load of ßPV types in eyebrow hair are associated with cSCC risk in OTRs, providing evidence that ßPV is associated with cSCC carcinogenesis and may present a target for future preventive strategies.

Longitudinal study of seroprevalence and serostability of the human polyomaviruses JCV and BKV in organ transplant recipients.

The human polyomaviruses BKV and JCV cause mostly subclinical infections in childhood. Systemical immunosuppression after organ transplantation can lead to reactivation of persistent polyomavirus infections which may cause rejection of the transplanted organ. BKV and JCV seroprevalence and serostability was measured in 441 European solid organ transplanted recipients. Baseline samples were collected on average 24 days post-transplantation and sera were then collected over an 18 months follow-up period on up to six different time points. The overall seroprevalence at baseline for BKV was 97% with very little change over time. Prevalence for JCV was 76% at baseline and increased to 80% at the end of follow-up. BKV seroprevalence was highest in the youngest age group (100%) and decreased with increasing age (92% in the oldest age group; P < 0.0001), while JCV increased with age (69% vs. 81%; P = 0.020). Antibody reactivities for both BKV and JCV increased significantly with time (P = 0.0002 and P < 0.0001, respectively). Among the 406 patients with several samples, 94% were stably seropositive for BKV and 1% remained seronegative during the follow-up. JCV antibody stability was somewhat lower: 67% remained stably seropositive and 13% seronegative. While seroprevalence of BKV and JCV decrease and increase with age, respectively, both polyomaviruses showed significant increasing antibody reactivity over time in organ transplanted recipients at the onset of immunosuppression.

Longitudinal study of seroprevalence and serostability of 34 human papillomavirus types in European organ transplant recipients.

Organ transplant recipients (OTR) are at increased risk of cutaneous squamous cell carcinoma, which may be related to reactivation of human papillomavirus (HPV) infections. Measurement of change in HPV antibodies after transplantation would help to explore this hypothesis. We measured antibodies to 34 HPV types on up to six occasions over 18 months in 441 OTRs from five European countries. At baseline (mean 24 days after transplantation), 80% of all OTRs were seropositive to at least one HPV type. The beta HPV genus had the highest seroprevalence (45%). For most HPV genera baseline seroprevalence peaked between 40 and 59 years old. Most OTRs retained their serostatus over time and antibody levels were stable. Seroprevalence in immunosuppressed OTRs is stable in the 18 months immediately after transplantation. Thus there is no short-term evidence that immunosuppression leads to new or reactivated skin infection with HPV sufficient to induce antibodies.

The risk of cancer is not increased in patients with multiple kidney transplantations.

BACKGROUND: The aim of this study was to investigate whether the number of transplantations, as a marker of the graft rejection status of the patient, is associated with an increased risk of malignancies. METHODS: In a cohort study, 1213 patients, receiving a kidney transplantation between 1966 and 1995 at the Leiden University Medical Center, were analyzed. All cutaneous squamous cell carcinoma and internal malignancies, which had developed between 1966 and 2007, were recorded. The influence of number of transplantations, age, sex and time on immunosuppression on the risk of squamous cell carcinoma and internal malignancies was investigated by time-dependent multivariate Cox's proportional hazard models. RESULTS: Of the 1213 kidney transplant recipients, 319 received a second kidney, 78 a third; 13 of them a fourth and 4 of them a fifth transplantation. After adjustment for potentially confounding factors, including age, sex and years on immunosuppressive therapy we did not detect an increased risk of cancer in patients with multiple transplantations. On the contrary, patients with three or more transplantations had a 1.6-fold decreased risk of squamous cell carcinomas and a 3.6-fold decreased risk of internal malignancies. CONCLUSION: We conclude that kidney transplant recipients with three or more transplantations do not have an increased risk of cutaneous squamous cell carcinoma and internal malignancies.

Kidney transplant recipients with cutaneous squamous cell carcinoma have an increased risk of internal malignancy.

This study aimed to investigate whether the occurrence of cutaneous squamous cell carcinomas (SCCs) is associated with an increased risk of internal malignancies (IMs) in kidney transplant recipients (KTRs). In a cohort study, all patients receiving kidney transplantation in Leiden, the Netherlands, between 1966 and 2006 were followed up. All malignancies that had developed between 1966 and 2007 were recorded. Time-dependent Cox regression analyses were used to calculate the association between the development of cutaneous SCCs and IMs. The incidence of IMs in the KTRs after transplantation was also compared with the general Dutch population by calculating standardized morbidity ratios (SMRs) and was matched for age, sex, and time period in which the malignancy had occurred. Among 1,800 KTRs, 176 (9.8%) developed cutaneous SCCs and 142 (7.9%) developed IMs after transplantation. In patients with prior cutaneous SCCs, the adjusted risk to develop IMs was 3.0 (1.9; 4.7). In KTRs without cutaneous SCCs, the risk of IM compared with the general population was hardly increased. KTRs with cutaneous SCCs have an increased risk to develop IMs, and this information can be used to identify KTRs who are at an increased risk for IMs.

Incidence of cancer in kidney-transplant recipients: a long-term cohort study in a single center.

In a long-term cohort study, we calculated cancer incidences and survival rates after the development of these cancers in kidney-transplant recipients. The cancer incidences were compared with those in the general population. The occurrence of cancer was recorded in all patients who received kidney transplantation between 1966 and 2006. The median follow-up time was more than 9 years with a maximum of almost 40 years. Altogether 327 (17%) of 1906 patients developed cancer after transplantation: 142 (7%) had non-cutaneous malignancies; 178 (9%) cutaneous squamous-cell carcinomas and 138 (7%) basal-cell carcinomas. The cumulative incidence of any cancer was 13%, 33% and 47% after 10, 20 and 30 years, respectively. The incidences of cancers of the oral cavity, stomach, female genital organs, kidney, thyroid gland, leukemias and lymphomas, and cutaneous squamous-cell carcinoma were significantly increased with a highest standardized morbidity ratio of 40 for cutaneous squamous-cell carcinomas. Survival rates after non-cutaneous malignancies were 57%, 43% and 36% and after non-melanocytic skin cancer 99%, 90% and 77% after 1, 3 and 5 years, respectively. The increased incidence of non-cutaneous malignancies after kidney transplantation is associated with a high mortality. Prevention of cancer after kidney transplantation should be a major focus of future research.

Increased risk of squamous-cell carcinoma in simultaneous pancreas kidney transplant recipients compared with kidney transplant recipients.

The purpose of this study was to ascertain the risk of non-melanocytic skin cancer (NMSC) in simultaneous pancreas kidney transplant recipients (SPKTRs) compared with kidney transplant recipients (KTRs) in relation to other potential risk factors of skin cancer. In a cohort study, 208 SPKTRs were compared with 1,111 KTRs who were transplanted during the same time period. The effects of age, sex, country of origin, time period after transplantation, HLA matching, immunosuppressive regimen, and rejection treatments on the risk of NMSC were investigated in multivariable Cox's proportional hazard models. In SPKTRs, the incidence of NMSC increased from 19 to 36%, respectively, 10 and 15 years after transplantation, which was significantly higher compared with that in KTRs (6 and 10%, respectively). After adjustment for age and sex, SPKTRs had a 6.2 (3.0-12.8) increased risk of squamous-cell carcinoma (SCC) compared with KTRs. An additional adjustment for maintenance immunosuppression decreased the hazard ratio to 3.1 (1.3, 7.2), which indicates partial confounding by the immunosuppressive regimen. Adjustment for induction and rejection therapy or HLA mismatching did not change the hazard ratio significantly. SPKTRs have an increased risk of SCC compared with KTRs, despite partial confounding by the immunosuppressive regimen.

Keratotic skin lesions and other risk factors are associated with skin cancer in organ-transplant recipients: a case-control study in The Netherlands, United Kingdom, Germany, France, and Italy.

This study examines the association of keratotic skin lesions with the development of skin cancer in 915 solid organ-transplant recipients in five European countries. In a hospital-based case-control study, cases with squamous- and basal-cell carcinoma were compared with controls without skin cancer. Questionnaires, scrutiny of medical charts, and skin examination were delivered according to a standardized protocol. Keratotic skin lesions and viral warts were counted on different body sites. Keratotic skin lesions were strongly associated with an increased risk of squamous-cell carcinoma, with adjusted odds ratios of 4.1 (2.4;7.0) and 12.1 (6.1;24) for 1-49 and 50 and more keratotic skin lesions compared with no lesions, respectively. Keratotic skin lesions were also associated with basal-cell carcinoma with adjusted odds ratios of 2.9 (1.7;4.9) and 4.0 (1.7;9.2) for 1-49 and 50 and more lesions, respectively. Lighter skin types and painful sunburns were also significantly associated with an increased risk of squamous- and basal-cell carcinoma. Keratotic skin lesions are strongly associated with skin cancer and are, thus, an important clinical criterion for identifying those organ-transplant recipients at an increased risk of skin cancers who should be offered more intensive skin surveillance.

Photodynamic therapy does not prevent cutaneous squamous-cell carcinoma in organ-transplant recipients: results of a randomized-controlled trial.

A randomized-controlled trial with paired observations was performed with 40 organ-transplant recipients to assess the preventive effect of photodynamic therapy (PDT) on the development of new squamous-cell carcinomas and to evaluate the effect of PDT on the number of keratotic skin lesions. The treatment area consisted of a randomly assigned forearm and the corresponding hand, whereas the other forearm and hand served as the control area. After the initial visit, follow-up visits were scheduled at 3-monthly intervals during 2 years. No statistically significant difference was found in the occurrence of new squamous-cell carcinomas between the treated and untreated arms: after 2 years of follow-up, we observed 15 squamous-cell carcinomas in nine out of 40 PDT-treated arms and 10 squamous-cell carcinomas in nine out of 40 control arms. The number of keratotic skin lesions increased in both arms, but was less pronounced in the PDT-treated arm. After 1 year of follow-up, a trend in favor of the PDT-treated arm was observed, but statistical significance was not reached. Nearly 80% of the patients reported mild to severe adverse effects consisting of pain and a burning sensation, immediately after the treatment. No long-term adverse events were noted. In conclusion, PDT does not appear to prevent the occurrence of new squamous-cell carcinomas in organ-transplant recipients, but to some degree, reduces the increase of keratotic skin lesions.