Morphoproteomics

Morphoproteomics is a novel medical discipline that combines histopathology, molecular and cellular biology and protein biochemistry to define the protein circuitry in diseased cells. In this way it should be possible to identify specific target for customized therapeutic intervention. One preclinical study on prostate cancer overlaps data from phosphorylation analysis with phospho-specific probes and morphological evaluation of cellular compartmentalization. The identification of peculiar localization of certain phospho-analytes demonstrated the efficacy of this kind of personalized studies and their clinical utility. Another study demonstrated the feasibility of this approach in neck and head squamous carcinoma, that often has poor prognosis and high mortality rate. In this case, mTOR pathway was analyzed in terms of potential clinical significance of its activated components. Rapamycin is an inhibitor of this pathway. The inhibitory effects on cellular growth were directly correlated with mTOR activation, and a paint of the block was obtained by immuno-hystochemistry. On conclusion, morphoproteomics gives details about the signal transduction by using the combination of several techniques. This approach has been positively evaluated in certain disease, like cancer where an alteration in protein circuitry often causes the disease. Other pathological model would be available soon to validate this approach also in other disease.

Acute gastroenteritis in children

Acute gastroenteritis affects numerous children at least one time in their infancy and childhood. This disease is usually due to viral infection and none treatment is suggested to treat the infection self. In contrast, more concerns are due to dehydration associated with vomiting and diarrhoea.
Dehydration is treated with oral or intravenous intake of fluid, but generally intravenous treatment is preferred because the high likelihood of emesis may reduce benefits. Anti- emetic drugs are not suggested for children for their important adverse effects. Anyway, several studies have been reported in literature that show the effectiveness of ondansetron, an antagonist of serotonin receptor. This drug has been initially developed to treat nausea following chemotherapy in cancer patients; up to date ondansetron seems also promising to care emesis in children during acute gastroenteritis. The major benefit is the reduction of vomiting episodes that make possible to use also oral rehydration treatment. Furthermore, duration of hospital staying or admission rate are also reduced after ondansetron treatment, showing a financial advantage in comparison to traditional approaches. In conclusion, these studies are very important to demonstrate and ascertain the effectiveness of drugs in children in well structured trials. It’s auspicial that other drugs will be analysed at the same manner.

The IntOGen interface

Sometimes, there is a gap between experimental biology and clinical medicine while a continuous interchange would be auspicial to well direct experiments and keep updated the therapies. An interesting tool has been developed at the Barcelona Medical Research Park (Spain).
IntOGen is a frame work that collects, integrates and manages data derived from genome- wide experiments on large scale projects such as the Cancer Genome Atlas and the International Cancer Genome Consortium. Scientists manually annotate all samples by using the International Classification of Disease for Oncology vocabulary, in terms of tumour topography and morphology. Furthermore, they apply statistical methods to identify the most relevant alterations, by analyzing multiple studies on the same kind of tumour. Finally, they consider the role of whole biological modules, such as a pathway, to demonstrate the involvement of a single gene altered. The website www.intogen.org is available for free and allows to know modules and genes important in cancer, share experiments and analyze data in the context of cancer. This interface has been built to fill the gap between medicine and molecular biology. Similar tools should be really useful not only for cancer but also for other kind of diseases, such as neurodegenerative disorders.

Hela cells, a story lasting 60 years.

Henrietta Lacks died in 1951 at the Johns Hopkins hospital in Baltimore because of an aggressive form of cervical cancer. She would be probably unknown now, if your cells hadn’t been extracted and cultured as HeLa cells. Scientists of every molecular biology and cellular lab know about these cells because they have used them at least one time or because they have studied their application on biology books. However, it’s very interesting the history about HeLa cells. Henrietta was 31 years old when she died and she had five children. She was the unwitting protagonist of the story, because on 1950s none informed consent was asked her. Doctor Gey and his wife had all scientific merits to make possible HeLa cell culture. They put these cells on Petri dishes; at that time they were performing a lot of experiments to try to culture human cell lines. HeLa cells were able to quickly grow in established conditions, differentially from other cells tested. Dr Gey sent his cells to many laboratories around the world and shared information about culture conditions and so on. This generosity allowed important scientific advances, especially in vaccination field because HeLa cells were firstly used to test and produce the Polio vaccine. Unfortunately, giving their ability to grow also in unfavorable conditions, HeLa cells became one of the most dangerous contaminant agents of other cell lines. The doubt that scientists were using HeLa cells in their experiments rather than breast, prostate or placental cells made necessary further analyses to figure out the true identity of cell lines used. Thus, after almost thirty years from her death, the Hopkins Hospital contacted Henrietta’s children and familiar to invite them donate some blood or tissue samples.
Genetic analyses and blood type were information required by scientists to complete the Henrietta’ profile and recognize HeLa cells from others. Even if the scientific purpose was correct, Henrietta’ family didn’t have all explanation needed to well understand physicians’ operations. This was only one of dark points from an ethical point of view present in this story. Fortunately after Henrietta’s experience, ethical question has acquired great importance in experimental medicine and now informed consent is required for every medical action. Another important issue of this story were the moral and legal questions that arose about the commercial value of something derived from human body. Who may have the copyright of HeLa cells? With these cells several billions of dollars have been gained by pharmaceutical companies, research institutes and so on, but Henrietta’s family haven’t had any benefit. But on other hand, what has been the role of Henrietta in whole story? She was just a poor mother who died too soon.

CXCR1 in breast cancer

Scientists from the University of Michigan and the INSERM (France) identified Cxcr1 gene as differentially expressed in cancer stem cells in comparison with other cancer cells in breast malignancies.
CXCR1 is the receptor for interleukin 8 and has been involved in tumour progression and metastasis in several kinds of cancers, such as prostate, glioma, ovarian and breast cancers. Furthermore, IL8 has been implicated in self renewal of stem cells in vitro. French researchers tested some small molecule inhibitors targeting CXCR1 or some antibody against this receptor in order to evaluate the effect on cellular behavior. These inhibitors directly acted versus cancer stem cells and at the same time indirectly induced cell death in bulk tumour. The promising result was that the whole cancer population was eliminated. A possible explanation for this larger effect could be the release of FAS ligand after inhibiting CXCR1 that determined apoptosis in all cancer cells. In animal model CXCR1 inhibitors reduced tumour mass and blocked metastasis, either alone or in combination with other drugs. Another possible approach to block cancer progression by acting on CXCR1 pathway is to interfere with IL8 production. Scientists from the University of Texas demonstrated that siRNA-IL8 reduced tumour weight in comparison to control in animal models. These studies seem really promising but further proof is necessary to validate CXCR1 pathway as a target for therapeutic intervention.

Monoclonal antibodies

Monoclonal antibodies are important tool in molecular biology, diagnostics and clinical studies. These protein are produced by a single cells isolated from immunized animals. Current protocol requires an immunization of an animal host, for instance rabbit or mouse; then spleen cells are selected in vitro and B cells are isolated. B cells from spleen are fused with tumoral mouse cells of mieloma in order to stabilize and make possible the B lymphocyte culture. Indeed, B cell culture is difficult to set up and maintain.
The hybridoma technology allows overcoming these difficulties because of genetic transformation of mieloma cells. Finally, hybridomas are serially diluted and the antibodies are obtained from cell hybridomas cultures derived from a single cell. Which are the advantages of monoclonal antibody in respect with polyclonal ones? Monoclonal antibodies are codified by the same gene and none point mutations are present to generate some difference into antibody population. Thus, the whole population is identical and specifically recognizes one antigen. Cross reactivity is reduced with monoclonal antibodies and the interaction between antigen and antibody is usually more stable. Furthermore, this technique is also really flexible because it’s virtually possible to create antibody versus each antigen, when it’s possible to immunize the host animal. Which are the applications for monoclonal antibody? Monoclonal antibodies are currently used in molecular biology and biochemistry laboratories for imaging, western blotting, immunoprecipitation and so on. A lot of protocols are based on antibody use. In diagnostics, monoclonal antibodies are used in ELISA dosage or in flow cytometric analyses, as well as infection detection or pregnancy diagnosis.

Clinical applications of monoclonal antibody are prevalently in oncology. In 1997, the first monoclonal antibody was approved for non- Hodgkin lymphoma treatment. Since this year, several antibodies have been optimized against breast cancer, leukaemia, colon cancer and recently lung cancer. Each antibody recognizes a tumoral antigen and specifically kills only the cells (tumoral) that present that molecule. Thus, adverse effects associated with the use of monoclonal antibodies are reduced if compared with traditional drugs. Based on their specificity, antibodies can be used to carry other useful drugs to cells. For instance, an antibody can be conjugated to radioactive compounds to be addressed to cancer cells. Furthermore, other drugs can be carried into the brain, giving the capability of monoclonal antibody to overcome the blood- brain barrier. Parkinson’s disease can be treated with this approach. Improvement of biochemical characteristics of monoclonal antibodies is one challenge for scientists for the next future. Indeed, it’s important to improve the delivery of monoclonal antibody into all districts of human body. The specificity will be a must if clinical or diagnostic applications are planned for the monoclonal antibody. Furthermore, cheaper technology must be optimized to allow large scale production. Research development in this field is really promising.

Novel function for IHD1 in glioma

Genetic mutations are usually identified in cancer. These alterations can cause loss of function, when one mutation interferes with protein functionality, or gain of function when a mutated protein over-works or is over-expressed in a wrong cellular district. In this case, a protein maintains the same function as a normal one. In some cases, mutation can generate novel functions in a protein.
This is the case of IDH1, the isocitrate dehydrogenase 1 that usually converts the isocitrate into alpha- ketoglutarate. When this protein is mutated, an overproduction of 2 hydroxyglutarate (2HG) is abundantly recovered into cells. 2HG is toxic for brain and its presence is correlated with cancer. X-ray diffraction demonstrated that the mutation at arginine 132 results in the formation of a distinct active site, different from those that catalyzes the normal reaction. So, a novel mutation has been identified and a novel function has been recognized. IDH1 could be considered a new target for therapeutic intervention. A selective and specific small molecule inhibitor could interact only with the mutated form, without interfering with the wild type protein and the glucose metabolism. This discover could hopefully help scientists to identify a treatment for glioma.

Novel mouse model for BRCA1 breast cancer

BRCA1 is one important marker for breast cancer; in particular BRCA1 is mutated in the invasive ductal cancer and in general is associated with lack of oestrogen receptor, progesterone receptor and Her/ ERB2, making useless current therapies for breast cancer. Giving the lethality of double mutant Brca1, none animal models were available up to date to study the role of this gene in tumorigenesis.

Everyone knows the importance to study cancer from its molecular basis. Scientists from the Netherlands Cancer Institute of Amsterdam proposed a conditional mutant in which tissue specific inactivation of Brca1 was performed by Cre Recombinase. This system allows to dispose on one new model that mimics several characteristics of human mammary tumour. Furthermore, numerous new inhibitors or useful molecules could be tested on this model in order to find out some drugs to care BRCA1 cancers. The work, published on the British Journal of Cancer, open new perspectives in breast cancer research. Indeed, the next generation of mouse models will regard resistance mechanisms and double mutant animals will be likely generated in order to highlight the correlation between lack of BRCA1 and the role of other proteins often mutated in breast cancer, such as TP53.

Direct to consumer genetic services

Large expansion of direct to consumer genetic services is observed world-wide. This service offer single gene tests as well as screening of customer’s DNA at various loci to identify some polymorphism associated with certain disease. For instance, in women with familiar past history of breast cancer can be monitored for BRCA1 and BRCA2 genes, that are recognized as elevated risk factors. Furthermore, some genes involved in Alzheimer’s disease can be tested. A study was performed to evaluate the consequences of knowing the genetic makeup and the possible disease encountered in the future: it seems that knowledge about genetics doesn’t influence life, for instance by generating depression or anxiety. More concerns are about the scientific value of these tests, the modality to perform them and the clinical utility. Different approaches have been done by government to regulate this modern phenomenon: for instance, in Germany these tests are forbidden and only physicians can suggest genetic test carried out in appropriate institutions. In United Kingdom it has been proposed a self- policing by industry. In the US regulation is up to Federal states and varies largely. First implication of the large diffusion of this kind of test is the increase of analysis and examination to confirm genetic information. Medical intervention can not be the direct consequence of the test, but more deepened talks with physicians and genetic counsellors should be preferred.

One odd and dramatic case in medicine

In 2007 a young Japanese woman gave birth to her child and after only one month died for leukaemia, ABL-BRC positive. During her pregnancy she didn’t know to be sick and also doctors didn’t diagnose the disease. After eleven months, also the baby showed a tumour into his cheek. Physicians excluded a possible sarcoma and diagnosed also in this case leukaemia, ABL-BCR positive, the same mother’s disease. Further analyses demonstrated that no genetic material from the father was present in baby’s tumour, this means that cancer cells derived only form the mother and escaped the foetal barrier into uterus. Scientist has suspected that some forms of cancers, such as leukaemia and melanoma that are prompt to metastasize, could move form mother to fetus and this unfortunate story gave the genetic proof. Cancer cells can escape our immune system and also foetal barrier, knowing molecular basis of this mechanism could help us to better understand cancer biology and find new strategies for therapeutic intervention. Fortunately, there are few possibilities that the case of Japanese woman can repeat and we can learn a lot from odd examples like this. A positive note to conclude this post: the baby was successfully cared and is still alive.