Theme-based Research Scheme 2016/17 (Sixth Round) Layman Summaries of Projects Funded

Theme 1: Promoting Good Health
Project Title: Plasma DNA as a Platform Technology for Cancer Detection
Project Coordinator: Prof Dennis Yuk-ming LO (CUHK)

Cancer is the top killer in Hong Kong. Approaches for the early detection of cancer hold promise for improving our ability to deal with this deadly disease. The analysis of tumour-derived DNA in plasma, referred generally as "liquid biopsies", is now a hotly researched topic in oncology. The project team is a world-leading group in the diagnostic applications of circulating nucleic acids in plasma. This project is designed as a continuation of the RGC Theme-Based Research Scheme (TRS) Project T12-404/11. Over the previous 5 years, the project team has developed a number of innovative genomewide approaches for detecting cancer-associated genomic, methylomic and transcriptomic aberrations in plasma. The project team has also undertaken a 20,000-persons prospective study for using plasma DNA for screening nasopharyngeal carcinoma (NPC). This study has provided extremely promising evidence that plasma DNA analysis does facilitate the early detection of cancer. In the present proposed project, the team aims to develop and refine the molecular methodologies in search for approaches that could achieve high sensitivity and specificity for the non-invasive detection of multiple cancer types. The research will build on foundation laid by the earlier project and will have an emphasis on the common malignancies in Hong Kong, namely hepatocellular carcinoma, colorectal carcinoma, NPC, prostate carcinoma, lymphoma and leukaemia.

Theme 1: Promoting Good Health
Project Title: Understanding Cancer Stemness in Liver Cancer - From Regulation to Translational Applications
Project Coordinator: Prof Irene Oi-lin NG (HKU)

Liver cancer (hepatocellular carcinoma, HCC) is a common malignancy worldwide and very prevalent in Southeast Asia and China including Hong Kong, due to the high prevalence of HBV infection. Its incidence is also one of the most rapidly increasing in the west. It is an aggressive cancer with high rate of recurrence even after surgical resection and frequent metastasis. In HCC, like other cancer types, a subset of cancer cells referred to as cancer stem cells (CSCs), or cancer propagating cells, is capable of self-renewal and maintaining tumor propagation. Notoriously, these cells are resistant to conventional chemo/radio-therapy and hence are responsible for sustaining tumor growth and recurrence. Therapies that can eradicate these cells may lead to cancer cures or control. We have identified a majority of the liver CSC markers in addition to those identified by others. We have provided solid evidence that the CSCs characterized by these markers have a functional phenotype of self-renewal and tumor propagating capabilities, and are highly tumorigenic, metastatic, and chemo- and radiation-resistant, features accounting for tumor recurrence. Of note, our compelling evidence also shows that antibodies selectively blocking the liver CSCs marked by these markers are efficacious in suppressing HCC tumor growth. This would require that we rethink the way we treat tumors, as our objective must not only focus on eliminating the rapidly dividing cells (the bulk tumor) that are the current targets of treatment, but should also aim at targeting the CSCs that fuel tumor growth. Thus, CSCs are considered a pivotal target for the eradication of cancers.

In this project, based on our and others' experience, we postulate that understanding the underlying mechanisms of liver cancer stemness and targeting the liver CSCs may provide an opportunity for novel treatment of HCC. Here, we aim to comprehensively identify the molecular pathways common to these different CSC subpopulations and examine the molecular mechanisms how these pathways may regulate CSCs, hence providing targets for selective blocking of CSC functions. We propose to also understand the lineage hierarchy and biology of these CSCs, which are at present unknown, using single cell transcriptome analysis. On the other hand, as cancer cells are maintained in a specialized niche microenvironment, delineating the tumor microenvironmental factors in the regulation of liver cancer stemness and CSCs will be strategic to identify new mechanisms. To this end, we aim to understand the infiltrating immune cells and stromal cells sustaining cancer stemness while tapping into relevance of immunotherapy targeting liver CSC. While we hope to dissect the underlying mechanism of liver cancer stemness, it is also our strong intention to translate the high-quality research findings from this project into pre-clinical trials to target liver CSCs for novel HCC treatments. During the course of the studies, we will continue to strive to train young scientists in research of cancer stem cells and cancer biology, as well as educate the general public.

Theme 1: Promoting Good Health
Project Title: Gastric Cancer Genomics and Beyond - Moving from Patient Samples to 3D Organoid Cultures for Integrative Genomics Analysis, Drug Sensitivity Assays, Cell Biological Studies and Animal Models
Project Coordinator: Prof Suet-yi LEUNG (HKU)

Gastric cancer (GC) is the 3rd leading cause of cancer death worldwide and has a high incidence in China and Hong Kong. High mortality rates highlight the urgent need for new treatment methods. With major funding from the pharmaceutical industry and our tissue bank of over 1000 patients with comprehensive pathological and long-term patient outcome data, we generated a large-scale, multidimensional GC genomics database that encompasses whole-genome/exome sequencing and comprehensive molecular profiling of over 400 local patients. This is amongst one of the world's largest scale studies to date, and on-going study has led to the identification of many new GC driver genes, as well as segregation of GC into several molecular subtypes, each with a unique combination of driver alterations (Nat Genet 2011&2014). The challenge lies in understanding how each unique combination contributes to cancer development and their reliance on survival pathways that could translate into therapeutic opportunities. To achieve this, we have been highly successful at generating 3D organoid cultures from patient normal and cancerous gastric epithelial cells that cover the diverse molecular subtypes, creating combination driver mutations using CRISPR, and performing drug sensitivity testing. Our team constitutes clinicians and scientists that span the boundaries of biology and bioinformatics, and the world's pioneering overseas labs, with a successful history of collaboration. We will 1) generate stepwise combination cancer driver alterations re-capitulating the different molecular subtypes from normal gastric organoids to characterize gene function and understand drug response; 2) study GC organoids carrying corresponding combination alterations for drug sensitivity testing; 3) identify vulnerability, signaling pathway deregulation, cancer stem cell phenotypes and drug resistance mechanisms using live cell imaging, systems biology approaches, and combinatorial functional genomic screens using our newly developed CombiGEM-CRISPR technology; 4) study the RHO signaling pathway deregulation we identified in diffuse type GCs including generating novel animal models; 5) develop new methods for integrative genomic analysis to identify novel cancer driver pathways and predict new drug targets that feedback to 1) for testing. Overall, this study is expected to accelerate therapeutic development of GC through deep biological insights on combination driver alterations, enabling genome-guided patient stratification and drug repositioning, identifying new driver genes and pathways for new drug development. The gastric organoid biobank will attract global funding and collaborations. This project will train future generations of scientists and clinicians that span biology, computational analysis and patient care, to compete at the forefront of the genomics era.

Theme 2: Developing a Sustainable Environment
Project Title: Diagnosis and Prognosis of Intensifying Eutrophication, Hypoxia and the Ecosystem Consequences around Hong Kong Waters: Coupled Physical-biogeochemical-pollution Studies
Project Coordinator: Prof Jianping GAN (HKUST)

Coastal eutrophication is caused by excessive nutrient loading which stimulates phytoplankton blooms when physical, chemical, and biological conditions are favorable. It may lead to harmful algal blooms (HABs) and hypoxia (or "dead zones"), both of which can threaten the ecosystem. Coastal eutrophication has been a global environmental issue for decades, yet its persistence reflects the scientific and socio-economic complexities involved in alleviating the problem.

The coastal waters around Hong Kong are also affected by persistent and increasing eutrophication. This deteriorating situation may increase the frequency of HABs, expand the area of hypoxic zones and lead to other ecosystem disruptions and worse of all, offset the environmental improvements achieved through the costly Harbour Area Treatment Scheme over the last decade.

Eutrophication/hypoxia in Hong Kong waters is essentially the ecosystem's response to the increasing nutrient discharge from the Pearl River and local sewage effluent. Meanwhile, the increasing discharge of organic pollutants also modulates the biogeochemical pathways and ecological consequences and further increases the severity of eutrophication/hypoxia. Highly variable oceanic currents transport the nutrients in the interactive river-estuary-shelf (RES) waters around Hong Kong. These waters and nutrients undergo complex coupled physical-biogeochemical processes and modulate eutrophication/hypoxia. To date, these key processes have not been investigated in a comprehensive manner in our RES waters or in similar ecosystems elsewhere in the world. Understanding the full spectrum of intrinsic coupled physical, biogeochemical, and pollution processes in eutrophication is crucial to predicting and mitigating the impacts of eutrophication, and it remains a huge scientific challenge regionally and globally.

By adopting a global and local perspective and conducting an interdisciplinary study with world-class methodology, we will investigate holistically the coupled physical-biological-chemical processes in this interactive RES system, and thereby develop tools for diagnosing and forecasting eutrophication/hypoxia. This represents the main intellectual merit of the proposed project. We will also develop a novel, state-of-the-art marine monitoring system by conducting interdisciplinary mapping and time-series measurements, from which we will further develop a novel coupled physical-biogeochemical-pollutant modelling system.

Our ultimate goal is to identify factors driving the increasing eutrophication and hypoxia, and to provide analytical tools and a scientifically-based strategy for stabilizing or even reversing the two and for ensuring the overall sustainability of the marine environment in Hong Kong. Four interlinked tasks will be performed to determine (1) the sources and sinks of nutrients and their biogeochemical controls, (2) ecosystem dynamics and biological controls, (3) pollutant and ecosystem impacts, and (4) physical controls, synthesis, and future trends in the RES waters.

Theme 2: Developing a Sustainable Environment
Project Title: Enhanced Separation and Sludge Refinery for Wastewater Treatment - Solving the Nexus of Pollution Control and Resource Recovery in Mega Cities
Project Coordinator: Prof Xiao-yan LI (HKU)

Many forms of development erode the environment, causing serious water pollution in Hong Kong, China and many other regions of the world. Most core wastewater treatment technologies were developed about half a century ago that are no longer capable of accommodating the fast population growth, industrialization and urbanization. A city like Hong Kong discharges more than 2 million tons of municipal wastewater every day. Removal of the pollutants in wastewater treatment is not only difficult and costly (~HK$3/ton) but also produces a large amount of sludge (~1 ton dewatered sludge per 1000 ton wastewater). Disposal of the sludge, and the food waste as well, is one of the most challenging and expensive environmental problems for large cities.

On the other hand, major pollutants (organics and nutrients) in wastewater are valuable resources that should be recovered instead of being degraded or wasted with the sludge. As a mega city with over 7 million people, Hong Kong is a typical urban environment needing such technological breakthroughs for its planned wastewater treatment upgrading from the current primary level to the secondary level. In this project, novel technologies, namely Enhanced Separation and Sludge Refinery (ESSR), will be developed for advanced wastewater treatment and food waste processing. The theme-based research includes the following programs:

(1) Chemically-enhanced Membrane Filtration (CeMF) replacing the conventional primary sedimentation, together with the side-stream Acidogenic sludge and food waste Co-Fermentation (sACF), for phosphate recovery and organic hydrolysis for use in production of organic acids and bio-plastics and in nitrogen removal by denitrification;
(2) Treatment of the waste sludge by thermal Sludge Hydrolysis followed by fungal Fermentation and Refinery (SHFR) for waste minimization and production of ethanol and bio-fibres, with the subsequent recovery of ethanol and ammonia from the solution;
(3) Integration and pilot demonstration of the novel CeMF-sACF and SHFR modules for advanced wastewater treatment with improved nutrient removal, energy saving, resource recovery and sludge reduction, together with the assessment of the effluent quality and its impact on the receiving aquatic ecosystem.

The proposed technological development will fundamentally transform wastewater treatment from an end-of-the-pipe purification to a resource-mining practice. Besides the design of new treatment plants, the novel processes can also be used as add-on modules to retrofit existing treatment facilities, achieving more sustainable water pollution control, resource recovery and sludge minimization for Hong Kong, China and elsewhere.

Theme 4: Advancing Emerging Research and Innovations Important to Hong Kong
Project Title: A Compact System for Terahertz Imaging and Spectroscopy
Project Coordinator: Prof Chi-hou CHAN (CityU)

According to the World Health Organization, an estimated 600 million people fall ill after consuming contaminated food, resulting in 420,00 deaths every year. Closer to home, much of Greater China has been plagued by food-safety fears due to the adulterated baby milk formula in mainland China a few years ago and the recent gutter oil scandal in Taiwan. Spate of tainted drugs containing industrial grade magnesium carbonate also rocked Taiwan in 2015. The agglomeration of food safety, nutrition, and food and drug security will put a strain on our healthcare system and impede our socioeconomic development.

Terahertz (THz) wave is in the electromagnetic spectrum between the conventional microwave and infrared regions with a wide range of applications. It is very sensitive to water molecules and capable of distinguishing intrinsic contrast between normal and cancerous tissues. THz spectroscopy can also differentiate plant oils and animal fats because their refractive indices and absorption rates vary differently with increasing frequency and temperature. THz technologies have also been used in the detection of melamine in milk powder, antibiotics in food matrices, pesticides in vegetables, and foreign objects in drugs and chocolates.

Researchers hoping to exploit this promising frequency regime must confront the enormous entry barriers, attributed to the cost of the testing equipment as well as the availability of THz sources with sufficient power. A low-cost, compact THz system for imaging and spectroscopy not only can accelerate THz research but also resolve more burning issues affecting our welfare. We have assembled an international, multidisciplinary team to develop highly-efficient THz sources and detectors, fast beam scanning devices for data acquisition, signal processing and imagery display for the proposed compact system. On the other hand, research on biomedical imaging, chemical spectroscopy, and imagery display could be simultaneously carried out using existing and currently built THz systems in our laboratory while the compact system is being developed. Our synergistic effort will bring advances in high-resolution biomedical imaging, inspection of materials for high value-added manufacturing and food safety, and will contribute to economies not only in Hong Kong but also Greater China and the rest of the world.

On completion of the project, our expected deliverables are:
- A compact THz system for imaging and spectroscopy;
- THz spectral libraries for chemical and biological compounds; and
- Fundamental science generated in the creation of the compact system as archived in highly reputable journals.

Theme 4: Advancing Emerging Research and Innovations Important to Hong Kong
Project Title: Learning and Assessment for Digital Citizenship
Project Coordinator: Prof Nancy Wai-ying LAW (HKU)

The project aims are to develop a research programme to (1) establish the key dimensions and indicators, as well as assessment instruments, for the establishment of developmental milestones for digital citizenship from childhood to young adulthood (age 7 to 22); (2) develop an online role play simulation game platform for fostering and assessing digital literacy and collaborative problem solving (two key digital citizenship competencies) for adolescents and young adults; (3) develop pedagogical theory and design principles for fostering digital citizenship based on massive empirical e-learning and assessment data; and (4) identify the family and school factors that contribute to the development of digital citizenship.

Mastery of the 3 Rs (Reading, wRiting and aRithmetic) has long been considered adequate for competent functioning in society. However, the advent of digital technology has not only accelerated the pace of social, economic, political and cultural changes, but also led to space-time compression. The emergence of social media and social networking technologies and increasing accessibility of digital mobile technologies have changed the worlds of work and leisure, and brought about new solutions and new challenges to our well-being. To ensure effective participation in the workplace, as well as personal and social well-being, citizens in the 21st century need to possess (i) digital literacy (technological fluency, media literacy and cyber-wellness); (ii) collaborative problem-solving ability (the holistic exercise of critical-thinking, creativity, collaboration and communication skills to address real-life problems through self-directed inquiry); (iii) self-regulation; and (iv) willingness to take risks. We assume that these four competencies are the hallmarks of digital citizenship.

Although there are research-informed, age-appropriate curricula, pedagogy and benchmarks for the 3Rs, there is a dearth of research on digital citizenship. This project will contribute to our basic understanding of developmental differences in the competencies that underlie digital citizenship and the family and school factors that contribute to their development. We will also deliver an online e-learning, data-collection and assessment platform for sustained research on digital citizenship. Collaborative problem solving (CPS), a key dimension of digital citizenship, is selected for focused study. An online serious game portal will be developed to serve the dual purpose of assessing CPS and investigating the effectiveness of different learning designs to support CPS development in diverse learners.