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Supplementary Figures 1-10 and Supplementary Tables 1 and 2

Several viruses are known to be associated with the development of certain cancers, including human papilloma virus (HPV), an established causative agent for a range of anogenital and head and neck cancers. However, the causality has been based on the presence of the virus, or its genetic material, in the sampled tumors. We have long wondered if viruses cause cancer via a "hit and run" mechanism such that they are no longer present in the resulting tumors. Here, we hypothesize that the absence of viral genes from the tumor does not necessarily exclude the viral aetiology. To test this, we used an HPV-driven oropharyngeal cancer (OPC) tumor model and CRISPR to delete the viral oncogene, E7. Indeed, the genetic removal of HPV E7 oncogene eliminates tumors in vivo. Remarkably, E7 deleted tumors recurred over time and develop new mutations not previously seen in HPV

Clinical trials and proof-of-concept pre-clinical work for different cancers has shown relatively positive outcomes and tumour killing when p53, a well-established tumour suppressor, levels and function is restored. Human papillomavirus (HPV) driven cancers encode the E6 oncoprotein that degrades p53 to allow HPV-driven carcinogenic process to proceed. Indeed, there have been several attempts in the past to revive p53 function in HPV driven cancers by pharmacological and genetic means to increase p53 bioavailability. Here, we employed a CRISPR activation (CRISPRa) approach to overcome HPV-mediated silencing of p53 by hyper expressing the p53 promoter. Hyperexpression of p53 led to HPV positive (+) cervical cancer cell killing and reduced cell proliferation. Increasing p53 bioavailability may have promising potential as a therapy for HPV driven cancers.

It has long been wondered if viruses cause cancer via a “hit-and-run” mechanism. We examined this in HPV-driven oropharyngeal cancer (OPC) using CRISPR to delete the viral oncogene, E7. Tumours regressed yet regrew over 3 months, despite the lack of E7, and had formed new mutations commonly found in HPV- OPC patient tumours. This is the first biological proof that HPV- OPCs can be formed via a hit-and-run HPV infection.

The major HPV oncogenes E6 and E7 are known for its notoriety in driving the carcinogenic process in human papilloma virus (HPV) driven cancers. It is well-established that the removal of E7 dampens HPV cancer cell growth and proliferation. This has made E7 one of the most attractive targets for HPV cancers. Seminal work from our laboratory employed in vivo CRISPR editing treatment to delete E7, resulting in the effective elimination of HPV positive (HPV+) cervical cancer tumours in vivo. We have also successfully delayed HPV+ tumour growth in vivo with aurora kinase (AURK) inhibitors, an effect which is strongly sensitized by the presence of E7 expression. Unlike our observations in cervical cancer cells, in vitro targeting of E6/E7 have only resulted in partial killing of HPV+ oropharyngeal cancer (OPC) cells. The effect of E7 removal on HPV+ OPC tumours in vivo have not been explored. In this study we investigated a staggered combination of aurora kinase inhibition, using alisertib, followed by CRISPR editing of E7, to determine if this would lead to better HPV+ OPC cell killing in vitro and in vivo. Remarkably, genetic deletion of E7 alone was sufficient to effectively eliminate established HPV+ OPC tumours in vivo suggesting that E7 is essential in the maintenance of HPV+ OPC cancers.

The major HPV oncogenes, E6 and E7, are known for its notoriety in driving the carcinogenic process in human papilloma virus (HPV) driven cancers. It is well-established that the removal of E7 dampens HPV cancer cell growth and proliferation. This has made E7 an attractive target for HPV cancers. Seminal work from our laboratory employed a CRISPR editing approach to delete E7 which resulted in the effective elimination of HPV

Plasmacytomas are monoclonal plasma cell tumors. They represent 3% of all orbital tumors. We present an unusual case of orbital plasmacytoma and advanced multiple myeloma where the sole presenting symptoms were those of diplopia and ptosis. Clinical examination revealed right hypertropia and variable left upper lid ptosis but no proptosis. The visual acuity and the rest of the ocular examination were normal in both eyes. An MRI scan of the brain and orbits revealed a frontal bone lesion consistent with a plasmacytoma. A bone marrow biopsy showed a light chain multiple myeloma with free lambda light chains. Although the literature reports diplopia and ptosis as being clinical features of orbital plasmacytomas, to our knowledge this is the first reported case where these symptoms were the only presenting features, despite widespread disease.

Accumulating evidence supports the concept that cancer stem cells (CSCs) are responsible for the tumor recurrence and metastasis, the two major causes of cancer-related death. Therefore, CSC-targeted cancer therapy is important for the future development of more effective and advanced cancer therapy. One of the approaches is to specifically silence oncogene expression in CSCs and inhibit their growth. The significance of this approach is its specificity and ability to avoid multi-drug resistance of CSCs. In this chapter, we will describe a method of silencing HPV oncogenes E6/E7 in human cervical CSCs using HeLa cells as a model system.

To investigate any potential effect of fibroid size and distribution on menstrual blood loss (MBL).Retrospective comparative study of 50 women with symptomatic fibroids who underwent uterine imaging and objective MBL measurement prior to uterine artery embolisation between 1999 and 2002.West of Scotland Gynaecology and radiology departments.Uterine imaging was by magnetic resonance imaging (MRI) in all but one case and MBL was performed using the alkaline haematin technique. Fibroid characteristics were assessed by an experienced radiologist unaware of the MBL measurements.Thirty-three (66%) women had objective menorrhagia with a MBL in excess of 80 ml per period. The commonest location of fibroids was intramural; these particular fibroids also had the largest diameter and the greatest uterine volume. There was a negative relationship between MBL and the diameter of the largest fibroid (r = -0.419, P0.01). All but two women (both of whom had subserosal fibroids alone) demonstrated distortion of the uterine cavity. All women with submucosal fibroids presented with menorrhagia.This study found that MBL correlated with neither fibroid size nor location. However, all the women with sub-mucosal fibroids had menorrhagia with a MBL greater than 80 ml.