Diagnosis and Prognosis Markers for Bone Cancers using Comparative
Biomedicine Approach
Bone tumors are devastating types of cancers usually detected at an advanced stage in both companion animals and human beings. Bone cancers i.e., osteosarcoma, Ewing sarcoma and chondrosarcma in both human and animals share common pathophysiological features enabling animal tumor studies to be directly translated to understand the human scenario. Especially the dog bone cancers have been proposed as a better model to understand the human aspect of disease, due to the similarities of disease origin and metastasis. Bone cancer early diagnosis and also monitoring the treatment progress (prognosis) is quite challenging and often performed using imaging and histopathology of invasive bone sampling. Especially in canines, chemotherapy is quite an expensive option and even the tumor may be not responsive due to advance stage of diagnosis. Also, the development of drug resistance has been reported in many human cases of osteosarcoma. Thus, there is a great need of devising novel early diagnosis and prognostic tools for bone cancers. In the cancer biology, EVs are gaining greater attention as a candidate for biomarkers discovery for early detection and also to monitor the cancer progression and treatment responsiveness due to the proteins, nucleic acids and other cargos carried by them. Thus, the present project aim to identify novel bone cancer biomarkers from the EVs present in blood and urine and also to use them as prognostic markers to monitor the progress and treatment responsiveness in both canine and human using advanced molecular tools with a multidisciplinary comparative medicine approach.
Project background
Primary bone cancer types like osteosarcoma, chondrosarcoma and Ewing sarcoma account for 1% of the diagnosed cancer among human and the incidence in the companion animals including cats and dogs are showing an increasing trend. Out of theses chondrosarcoma is the rarest form. In human and animals osteosarcoma is extremely malignant and show high degree of metastasis to lungs in both human and canine patients. Moreover, there are not many early diagnostic tools available to detect most of the bone cancer types, thus, usually the cases are being detected with metastasis and at an advanced stage of the disease. The treatments of choice are mainly the surgical resection/ amputations and multi-drug chemotherapy. However, most of the cases have very poor prognosis and if metastasis has occurred usually survival rates are very low and multi drug resistance also becoming a concern in the limited options available. However, with recent medical advancements use of more adjuvant and neoadjuvant chemotherapy in combination with surgery has helped to improve the survival rates among the various types of the bone caners. Osteosarcomas are the most common form (about 80%) in both human and companion animals like dogs. In dogs, mostly large breeds are affected and mostly the diagnosis happen when visible clinical signs are presented and then confirmation with radiography and bone biopsy based osteo-cytological examinations are carried out. In human, osteosarcoma most often occurs in children and adolescents. It typically develops in the metaphysis of long bones, specifically the distal femur, proximal tibia, and proximal humerus and usually it metastasis to the lungs. The Erwin sarcoma usually develops in the pelvis, ribs and scapula and chondrosarcoma usually occur in the cartilage containing bones and are commonly seen in adults. All three types can be metastatic to lungs and except chrodrosarcoma, the other two types can spread to any other bone tissue types. The origin of most of the bone cancer types is still is a mystery , however, empirical evidence is pointing out that primitive mesenchymal cells may produce the malignant osteomatric during the differentiation process classically in the long bone types. Moreover, the tendency of bone cancer metastasis to lungs is also a trivial question for researchers. Diagnosis of bone cancers is often bit late radiography, imaging and histopathology are routinely used in the diagnosis of bone tumors. However, there are a number of diagnostic issues when interprets bone biopsy samples, such as distinguishing reactive bone from tumor bone, fracture callus and tumor bone, different benign fibro-osseous lesions, and typing of bone sarcoma. Thus, common diagnostic decision for radiologists and pathologists is the question of whether the primary disease process is inflammatory (reactive) or neoplastic. Then if the process is neoplastic, deciding whether the tumor is benign or malignant. Moreover, especially in dogs chemotherapy is an expensive option for many owners thus, if typing and metastasis potential can be predicted at the diagnostic stage there will be lot of assistance to the veterinarian to decide whether euthanasia is an option rather than costly and painful chemotherapy for dogs. In human skeletal bones can be the recipient of metastasis from other malignancies, like breast, lung, renal, prostate, and thyroid cancers. Moreover, bone marrow can be the nidus of malignancy in blood cancer types like multiple myeloma, lymphoma, and leukemia. In all these the malignant cells are not of bony origin thus definitive differential diagnosis also essential in clinical decision-making process. Therefore, there is a pressing need of identifying precise, alternative and early diagnostic methods for bone cancer and also a niche is available for better prognostic biomarkers discovery for bone cancer too.
EVs have recently gained momentum in science as a as a novel mechanism of intercellular communication. Especially in the cancer biology, accumulating evidence shows that EVs play important roles in tumor metastasis (establishment of a premetastatic niche) promotion of angiogenesis, overcoming the blood-brain barrier, induction of drug resistance, and formation of the heterogeneity of cancer-associated fibroblasts. Furthermore, EVs carry a variety of molecules, such as proteins and nucleic acids, that reflect the phenotype of their parental cells or the origin and they may contain molecules which could be the earliest responses in the malignant transformation process of the cells including bone cells. In addition, as EVs are stably circulated in almost all kinds of body fluids, they also have great potential as useful disease biomarkers as well. As a component of liquid biopsy, they have been repeatedly reported to be stably detectable in various kinds of body fluids, such as blood, saliva, urine, bronchoalveolar fluid, breast milk, semen, uterine and follicular fluids etc. More importantly, they reflect the current disease state by carrying specific molecules from parental cells, such as proteins, miRNAs, mRNAs, long noncoding RNAs (lncRNAs), and lipids. Thus, targeting the cargo of EVs allows us to assess crucial molecular information about disease pathophysiology, progression of the disease and also response to the treatments. Furthermore, because of the minimally invasive nature of the sampling process and accessibility, EVs are suitable for sequential sampling for monitoring purpose. Therefore, EVs have great potential as clinically useful biomarkers that can provide multiple, minimally invasive snapshots of disease status especially in bone cancer where early diagnosis is still quite challenging.
In this project we aim to explore the possibility of utilizing blood and urine borne EVs as early biomarkers for bone cancer with the further aim of using them in disease monitoring and prognosis in both human and dogs using comparative biomedicine approach. Since canine models have been proved to be superior over the other known bone cancer models, the knowledge generated from bog bone cancer study will immensely help in understanding and translating the techniques for human bone cancer research (Figure 8).
Primary bone cancer types like osteosarcoma, chondrosarcoma and Ewing sarcoma account for 1% of the diagnosed cancer among human and the incidence in the companion animals including cats and dogs are showing an increasing trend. Out of theses chondrosarcoma is the rarest form. In human and animals osteosarcoma is extremely malignant and show high degree of metastasis to lungs in both human and canine patients. Moreover, there are not many early diagnostic tools available to detect most of the bone cancer types, thus, usually the cases are being detected with metastasis and at an advanced stage of the disease. The treatments of choice are mainly the surgical resection/ amputations and multi-drug chemotherapy. However, most of the cases have very poor prognosis and if metastasis has occurred usually survival rates are very low and multi drug resistance also becoming a concern in the limited options available. However, with recent medical advancements use of more adjuvant and neoadjuvant chemotherapy in combination with surgery has helped to improve the survival rates among the various types of the bone caners. Osteosarcomas are the most common form (about 80%) in both human and companion animals like dogs. In dogs, mostly large breeds are affected and mostly the diagnosis happen when visible clinical signs are presented and then confirmation with radiography and bone biopsy based osteo-cytological examinations are carried out. In human, osteosarcoma most often occurs in children and adolescents. It typically develops in the metaphysis of long bones, specifically the distal femur, proximal tibia, and proximal humerus and usually it metastasis to the lungs. The Erwin sarcoma usually develops in the pelvis, ribs and scapula and chondrosarcoma usually occur in the cartilage containing bones and are commonly seen in adults. All three types can be metastatic to lungs and except chrodrosarcoma, the other two types can spread to any other bone tissue types. The origin of most of the bone cancer types is still is a mystery , however, empirical evidence is pointing out that primitive mesenchymal cells may produce the malignant osteomatric during the differentiation process classically in the long bone types. Moreover, the tendency of bone cancer metastasis to lungs is also a trivial question for researchers. Diagnosis of bone cancers is often bit late radiography, imaging and histopathology are routinely used in the diagnosis of bone tumors. However, there are a number of diagnostic issues when interprets bone biopsy samples, such as distinguishing reactive bone from tumor bone, fracture callus and tumor bone, different benign fibro-osseous lesions, and typing of bone sarcoma. Thus, common diagnostic decision for radiologists and pathologists is the question of whether the primary disease process is inflammatory (reactive) or neoplastic. Then if the process is neoplastic, deciding whether the tumor is benign or malignant. Moreover, especially in dogs chemotherapy is an expensive option for many owners thus, if typing and metastasis potential can be predicted at the diagnostic stage there will be lot of assistance to the veterinarian to decide whether euthanasia is an option rather than costly and painful chemotherapy for dogs. In human skeletal bones can be the recipient of metastasis from other malignancies, like breast, lung, renal, prostate, and thyroid cancers. Moreover, bone marrow can be the nidus of malignancy in blood cancer types like multiple myeloma, lymphoma, and leukemia. In all these the malignant cells are not of bony origin thus definitive differential diagnosis also essential in clinical decision-making process. Therefore, there is a pressing need of identifying precise, alternative and early diagnostic methods for bone cancer and also a niche is available for better prognostic biomarkers discovery for bone cancer too.
EVs have recently gained momentum in science as a as a novel mechanism of intercellular communication. Especially in the cancer biology, accumulating evidence shows that EVs play important roles in tumor metastasis (establishment of a premetastatic niche) promotion of angiogenesis, overcoming the blood-brain barrier, induction of drug resistance, and formation of the heterogeneity of cancer-associated fibroblasts. Furthermore, EVs carry a variety of molecules, such as proteins and nucleic acids, that reflect the phenotype of their parental cells or the origin and they may contain molecules which could be the earliest responses in the malignant transformation process of the cells including bone cells. In addition, as EVs are stably circulated in almost all kinds of body fluids, they also have great potential as useful disease biomarkers as well. As a component of liquid biopsy, they have been repeatedly reported to be stably detectable in various kinds of body fluids, such as blood, saliva, urine, bronchoalveolar fluid, breast milk, semen, uterine and follicular fluids etc. More importantly, they reflect the current disease state by carrying specific molecules from parental cells, such as proteins, miRNAs, mRNAs, long noncoding RNAs (lncRNAs), and lipids. Thus, targeting the cargo of EVs allows us to assess crucial molecular information about disease pathophysiology, progression of the disease and also response to the treatments. Furthermore, because of the minimally invasive nature of the sampling process and accessibility, EVs are suitable for sequential sampling for monitoring purpose. Therefore, EVs have great potential as clinically useful biomarkers that can provide multiple, minimally invasive snapshots of disease status especially in bone cancer where early diagnosis is still quite challenging.
In this project we aim to explore the possibility of utilizing blood and urine borne EVs as early biomarkers for bone cancer with the further aim of using them in disease monitoring and prognosis in both human and dogs using comparative biomedicine approach. Since canine models have been proved to be superior over the other known bone cancer models, the knowledge generated from bog bone cancer study will immensely help in understanding and translating the techniques for human bone cancer research (Figure 8).
Project objectives
- Development of EVs isolation and characterization protocols for canine and human bone tumor /cancer patients’ from less or non-invasive samples.
- Exploration of diagnostic, prognostic and metastasis related biomarkers discovery from bone cancer / tumor patients derived EVs.
Integration with EMU expertise, research strategy and development
The proposed research work will be initially started with the dog patients since the Institute of Veterinary Medicine and Animal Science (IVMAS) of the EMU has well trained clinical staff and infrastructure in the clinics to select, recruit and sample the dogs. Then the collected biopsies will be processed by the scientists in the COMBOVET ERA chair to extract EVs and further subjected them for different proteomics and genomics analysis of the cargo/ This way the planned work will create a synergy between the clinical and basic science platforms to work towards a common goal. Furthermore, the pathology section of the IVMAS/EMU will come onboard performing the histopathological work related to the samples. Thus, the proposed work will be a multi-disciplinary and multi-dimensional research work aiming to uplift the research outcomes of the EMU to a next level using comparative biomedical approach. Furthermore, for the human sampling we will collaborate with the clinicians in the Hospital of the University of Tartu extending the collaborations beyond the EMU which is one of research and development the development goals of EMU.
The proposed research work will be initially started with the dog patients since the Institute of Veterinary Medicine and Animal Science (IVMAS) of the EMU has well trained clinical staff and infrastructure in the clinics to select, recruit and sample the dogs. Then the collected biopsies will be processed by the scientists in the COMBOVET ERA chair to extract EVs and further subjected them for different proteomics and genomics analysis of the cargo/ This way the planned work will create a synergy between the clinical and basic science platforms to work towards a common goal. Furthermore, the pathology section of the IVMAS/EMU will come onboard performing the histopathological work related to the samples. Thus, the proposed work will be a multi-disciplinary and multi-dimensional research work aiming to uplift the research outcomes of the EMU to a next level using comparative biomedical approach. Furthermore, for the human sampling we will collaborate with the clinicians in the Hospital of the University of Tartu extending the collaborations beyond the EMU which is one of research and development the development goals of EMU.
Project team
Suranga Kodithuwakku
Institute of Veterinary Medicine and Animal Sciences Investigator |
Alireza Fazeli
Institute of Veterinary Medicine and Animal Sciences Investigator |
Aleksandr Semjonov
Institute of Veterinary Medicine and Animal Sciences Investigator |
Aare Martson
Institute of Veterinary Medicine and Animal Sciences Investigator |