A  few  years  ago,  Javier  Benítez, director  of  the Human Genetics Group at the CNIO,  received a  call  from Pablo  García  Pavía,  from  the  Cardiology  Unit  of  the  Puerta  de  Hierro University Hospital. This cardiologist was treating two brothers with a rare form of cancer, cardiac angiosarcoma (CAS). Could the experts in genetics do  something?  “At  that  time  we  tried  a  few  ideas,  but  unsuccessfully,”says Benítez. We have had to wait for modern genome analysis techniques to  discover  the  brothers’  genetic  problem.  The  finding  opens  a  way  to identify  CAS  families  who  are  carriers  of  a  mutation  in  the  gene
responsible  for  the disease. Family members  could  then  benefit  from an early diagnosis and the appropriate treatment.

Researchers  in  Benítez’s  group  recently  revaluated  the  case  of  the
brothers  with  CAS.  After  sequencing  their  exome  —  the  part  of  the
genome  that  is  translated  into  protein  and  therefore  the  one  that  most
influences the state of the organism, they  found that the cause of the
illness was a mutation in a gene called POT1.

The  identification  of  this  gene  led  them  directly  to  another  CNIO  group,
the  Telomere  and  Telomerase  Group,  headed  by  María  Blasco.  POT1  is
one of the proteins that comprise the protective shield around telomeres
—  the  structures  that  protect  the  tips  of  chromosomes —  and  it  has
recently  been  identified  as  responsible  for  other  forms  of  hereditary
cancer: melanoma  and  familial  glioma.  Blasco’s  group  is  not  only  one  of
the leading groups in  the field of  telomeres, but has also participated —
together with the groups headed by Carlos López-Otín and Elías Campo —
in  the  first  description  of  the mutation  of  this  gene  in  human  cancer
(chronic lymphocytic leukaemia).

Cardiac  angiosarcoma  is  a  rare  but  malignant  disease.  In  the  case  of
hereditary  CAS,  the  median  survival  expectancy  is  only  four  months
because  the  disease  is  diagnosed  at  an  advanced  stage.  Until  now,  no
related gene has been identified.

CNIO researchers also observed that hereditary CAS occurs in families with
a  high  incidence  of  other  types  of  cancer.  This  is  similar  to  what  is
observed in people affected by the so-called Li-Fraumeni syndrome, which
is caused by a mutation in the tumour suppressor gene — nicknamed the
genome  guardian — P53.  However,  POT1, but  not  P53, was  found
mutated in the families affected by CAS.

The  discovery  of  the  new  mutation  proved  to  be  even  more  significant
from  a  clinical  perspective,  given  that  it  identified  carriers  at  risk  of
developing cardiac angiosarcoma and possibly other tumours.

As  Benítez  explained,  “in  the  past,  we  simply  didn’t  have  anything  that
could  help  in  identifying  these  people  at  risk,  because  there  were  no
markers  for  familial  CAS  or  for  families  with  a  syndrome  similar  to  Li-
Fraumeni  without  P53  mutations.  This  study  uncovers  one  of  the  genes
that explains the high incidence of cancer in some of them.”

“The translation of these results into the clinic is immediate,” says Blasco.
“In fact, we are already helping families that carry this mutation.” CNIO

Unravelling the genetic sequences of cancer that has spread to the brain could offer unexpected targets for effective treatment, according to new research.

Researchers say that they found that the original, or primary, cancer in a patient’s body may have important differences at a genetic level from cancer that has spread to the patient’s brain (brain metastases). This insight could suggest new lines of treatment.

Dr Priscilla Brastianos, MD, a neuro-oncologist and Director of the Brain Metastasis Program at Massachusetts General Hospital, Boston, USA, said: “Brain metastases are a devastating complication of cancer. Approximately eight to ten percent of cancer patients will develop brain metastases, and treatment options are limited. Even where treatment is successfully controlling cancer elsewhere in the body, brain metastases often grow rapidly.”

Dr Brastianos and her colleagues studied tissue samples from 104 adults with cancer. In collaboration with Dr Scott Carter and Dr Gad Getz at the Broad Institute, Cambridge, USA, they analysed the genetics of biopsies taken from the primary tumour, brain metastases and normal tissues in each adult. For 20 patients, they also had access to metastases elsewhere in the body.

Brain metastases often manifest years after the primary tumour. Before this study was carried out, the extent to which the genetic profiles of brain metastases differ from that of the primary was unknown.

The researchers found that, in every patient, the brain metastasis and primary tumour shared some of their genetics, but there were also key differences. In 56% of patients, genetic alterations that potentially could be targeted with drugs were found in the brain metastasis but not in the primary tumour.

“We found genetic alterations in brain metastases that could affect treatment decisions in more than half of the patients in our study,” Dr Brastianos will say. “We could not detect these genetic alterations in the biopsy of the primary tumour. This means that when we rely on analysis of a primary tumour we may miss mutations in the brain metastases that we could potentially target and treat effectively with drugs.”

This study also found that if a patient had more than one brain metastasis, each was genetically similar.

To date, scientists have had limited understanding of how cancers change genetically, or evolve, as they spread from the primary tumour. The researchers used their findings to map the evolution of a cancer through a patient’s body, and draw up a so-called phylogenetic tree for each patient to demonstrate how the cancer had spread and where each metastasis had come from.

They concluded that brain metastases and the primary tumour share a common genetic ancestor. Once a cancer cell, or clone, has moved from the primary site to the brain, it continues to develop and amass genetic mutations. The genetic similarity of the brain metastases in individual patients suggests that each brain metastasis has developed from a single clone entering the brain.

The genetic changes in brain metastases are independent of any occurring at the same time in the primary tumour, and in metastases elsewhere in the body, the researchers said.

Characterisation of the genetics of a patient’s primary cancer can be used to optimise treatment decisions, so that drugs that target specific mutations in the cancer can be chosen. However, brain metastases are not routinely biopsied and analysed. ECC 2015

Ortho Clinical Diagnostics is emerging stronger than ever since becoming an independent company in 2014, when it was purchased by The Carlyle Group, and has made tremendous progress in developing a range of new assays. Company researchers presented data from five assays currently under development at this year’s American Association for Clinical Chemistry (AACC) meeting.

“We are investing in our business to better serve the modern clinical lab with state-of-the-art solutions,” said Ted Farrell, Vice President Business Field Assays. “We continue to enhance the quality of our products and expand our new product development pipeline for our Clinical Laboratory business.”

The assays presented at AACC address a range of important areas for clinical lab testing including HIV detection and cardiac event monitoring. Following is a quick overview:

• Ortho Clinical Diagnostics is developing a fourth generation assay to detect both HIV 1 antigen genotypes and HIV 1 & 2 antibody subgroups for use on its random access VITROS® systems. The assay demonstrated seroconversion sensitivity consistent with a commercially available fourth generation assay and was more sensitive than a third generation assay.
• A rapid, fully automated, high sensitivity assay is under development for the measurement of cardiac troponin I (cTnl) and is designed to be more analytically sensitive than contemporary cTnl and cTnT assays.
• Preliminary performance data showed that OCD’s prototype VITROS^® Insulin Assay has excellent precision, cross-reactivity with pro-insulin and c-peptide as well as good correlation with two methods that are already commercially available.
• The current VITROS^® Cl- Slide is FDA cleared for use with serum and plasma, but not in urine.Testing of urine samples using the current calibration and the proper testing protocol for plasma and serum resulted in impressive performance, reproducibility and linearity.

Ortho Clinical Diagnostics is focused on bringing targeted solutions like these to its clinical laboratory customers aimed at addressing unmet clinical needs and driving improvements to quality care. It continues to press the boundaries of what’s possible in its quest for new and better assays.  

DIAsource ImmunoAssays has received China FDA clearance for its 25OH Vitamin D ELISA assay based on proprietary (Patented) monoclonal antibodies. The test is successfully launched in the Chinese market in collaboration with a local Chinese distributor and its sub-distributors. The assay which has the CE mark and is FDA cleared, is characterized by a very simple protocol with an extremely efficient pretreatment solution directly in the ELISA well. Its extreme user-friendliness makes it a popular assay among manual ELISA users as well as in laboratories that need high throughput using open ELISA instruments. The company provides validated protocols for the most common ELISA automates in the market. This assay is one of the various assays for determination of Vitamin D (25OH Vitamin D, 1,25 (OH)2 Vitamin D and free 25OH Vitamin D) that DIAsource has available (IVD and RUO versions) in its product portfolio either in an ELISA and/or RIA format.

Rheonix Inc., a developer of fully automated molecular diagnostics solutions, has been granted patent 9,132,398, “Integrated Microfluidic Device and Methods,” for the Rheonix CARD^® cartridge, which enables assays to be performed on the company’s EncompassMDx^® and Encompass Optimum™ instruments. The CARD, which stands for Chemistry and Reagent Device, will make molecular diagnostics simpler and easier to perform through an innovative and functional design that delivers a fully automated molecular assay at a fraction of the cost of other options. All assay steps are performed within the fully enclosed cartridge, thus eliminating the potential for contamination, reducing user error, and streamlining workflow.
The CARD’s design enables adoption of advanced molecular technology by laboratories of all types, from small community hospital labs to highly complex, centralized laboratories. The design also facilitates implementation across a wide range of market opportunities, including next-generation sequencing (NGS) sample prep, research-use-only testing, food and beverage industry applications, and in vitro diagnostics.  
The ’398 patent allows researchers and clinicians to quickly, easily, and cost-effectively run several samples through a fully integrated and automated nucleic acid amplification test, from raw sample input through detection, with no user intervention. Each CARD allows for simultaneous testing of four different samples and can handle a broad range of sample types, such as fresh tissue, urine, whole blood, serum, saliva, swabs, and formalin-fixed, paraffin-embedded (FFPE) tissue. The Rheonix CARD performs multiple molecular techniques, including sample preparation, such as chemical and enzymatic lysis and DNA purification; amplification, such as endpoint polymerase chain reaction (PCR), reverse transcriptase PCR, and quantitative PCR; and detection on a low-density microarray or lateral flow strip.
“The ’398 patent recognizes the groundbreaking achievement we have reached with the Rheonix CARD. From the device’s hardware to its process, it will help make molecular diagnostics a reality in laboratories worldwide,” said Tony Eisenhut, president of Rheonix. “With the lowest cost of ownership of any molecular platform, the patent confirms the novelty of the Rheonix approach to molecular diagnostics. Where other systems have traditionally emphasized either multiplex or throughput, Rheonix has designed single-use cartridges that do both and can perform sophisticated functions with a simple design. This lowers laboratory costs by eliminating waste in time, equipment and consumables, and reduces the amount of highly skilled labour. Rheonix is helping bring powerful molecular tools to laboratories that could not previously afford to purchase or run them.”
The dual-layer design of the Rheonix CARD automatically manipulates reagents internally with its active fluidic network of pumps, valves and channels. The upper surface of the CARD contains reservoirs that hold reagents used in the extraction, purification, amplification and detection process and any resulting liquid waste. The channels and pumps located on the lower surface of the CARD are used to transport and mix reagents and move wastes into the reservoirs on the top. By actively pumping fluids from reservoir to reservoir within the CARD, molecular diagnostic tests can be performed automatically.