Laboratory of Translational Hematology and Chemogenomics

Scientific Supervisor: Prof. Giovanni Roti

For Info and Contacts:

Prof. Giovanni Roti, 0521-702200, giovanni.roti@unipr.it

Administrative: Dott.ssa Ester Regazzi, regazziester@gmail.com

Affiliating Staff:

- Prof. Giovanni Roti (PI), giovanni.roti@unipr.it

- Dott.sa Anna Montanaro(RTD-A), anna.montanaro@unipr.it

- Dr. Luca Pagliaro (RTD-A), luca.pagliaro@unipr.it

- Dr. sa Federica Vento (PostDoc Fellow), federica.vento@unipr.it

- Dr. sa Elisa Simoncini (PostDoc Fellow), elisa.simoncini@unipr.it

- Dr. Roberto Rosati (Computational Biologist), roberto.rosati@unipr.it

- Dr. sa Mariateresa Giaimo (PostDoc Fellow), mariateresa.giaimo@unipr.it

- Dr. Gregorio Monica (PhD Student), gregorio.monica@unipr.it

- Dr.sa Marika Malchionna (PhD Student), marika.malchionna@unipr.it

- Dr. Andrea Gherli (Fellow), andrea.gherli@unipr.it

- Dr. sa Raffaella Zamponi (Lab manager), raffaella.zamponi@unipr.it

- Dr. sa Ester Regazzi (Administrative), regazziester@gmail.com

The Translational Hematology and Chemogenomics (THEC) laboratory of the Department of Medicine and Surgery is located in Pavilion 15, Floor 1, V. Gramsci 14, 43126 Parma.

The Translational Hematology and Chemogenomics (THEC) laboratory is dedicated to the study of cancer Biology, with a specific focus on hematologic malignancies. The goal is to develop innovative therapies, mainly through multi-omics approaches. The research projects have translational implications and involve the collaboration of A.O.U. Parma Hematology. Specifically, THEC uses a combination of cell lines, primary samples, and patient-derived mouse models (PDX) and makes use of screening of "small molecules," genetic approaches (sh/sgRNA, CRISPR/Cas9), cell Biology and genomics (RNAseq, ChIP-seq, NGS) to identify new targets and therapeutic strategies.
One of the main lines of research involves the study of the pathway of Notch1, a transcription factor frequently mutated in acute T-cell lymphoblastic leukemias (T-ALL) and in a group of chronic lymphoproliferative diseases (CLL, MCL), with the aim of identifying new target therapies. In fact, the group has recently characterized a SERCA inhibitor (CAD204520), which, by indirectly modulating the Notch1 pathway, reduces the off-target effects typical of other SERCA inhibitors, while maintaining antileukemic properties in vitro and in vivo.
THEC is also involved in the study of other determinant transcription factors in hematologic malignancies. Although transcription factors represent an ideal target in molecular therapy, they have often been ignored in drug development, in part because of the difficulty of developing high-throughput screening for the identification of specific modulators and, in addition, because of the Chemistry inaccessibility of such molecules. Through the use of chemistry compound libraries and the application of "forward" and "reverse" chemogenomic screening approaches, THEC has characterized mechanisms of action and identified novel target therapies for undraggable transcription factors, such as MECOM1 in acute myeloid leukemia (AML), EHTM2 and WNK1 in T-ALL.
Finally, the laboratory has initiated the establishment of a national network for the use of a drug response profiling (DRP) platform, with the goal of integrating drug sensitivity profiles with genomic data and identifying new therapeutic options. This approach allows supporting clinical decisions with timeframes compatible with the care pathway and represents a promising tool to prospectively identify the most appropriate treatment strategies and achieve deeper therapeutic responses, contributing to the development of functional precision medicine in hematologic malignancies.

 

Techniques in use:

The most innovative methodologies involve drug profiling for the identification of personalized therapies. We have developed a pipeline that allows the processing of cells derived from bone marrow/peripheral blood of patients with libraries containing hundreds of different molecules. The process is highly automated and enables high-throughput experiments, with minimized human error, extreme repeatability, and time and material savings. The system's speed of execution allows it to work on freshly taken patient samples and to obtain translational results in a short time. THEC also performs synergy studies between combinations of multiple drugs. In vitro drug efficacy validation can be performed with high-throughput flow cytometry studies, which, in addition to monitoring drug response in terms of cell viability, provides immunophenotypic analysis and functional target discovery studies. The laboratory also uses innovative genome editing techniques (e.g., CRISPR-Cas9) to recapitulate and validate the Chemistry inhibition phenotype. Finally, with direct access to sequencing facilities, it performs NGS diagnostic evaluations in patients with hematologic diseases. This makes it possible to increase the complexity of research work and to identify more and more comprehensively the genetic and molecular mechanisms responsible for drug resistance.

Tecan D300e Digital Dispenser. Using HP Direct Digital Dispensing technology, it provides liquid dispensing directly onto the dispensing plate, without contact with it, from picoliters to microliters, saving time and minimizing sample consumption. It offers a simple method for generating dose-response curves, multi-drug synergy experiments, enzyme profiles, and setting up PCR reactions

Attune NxT cytofluorometer. Compact analytical system that enables evaluation of cytofluorimetric characteristics, ensuring consistent and reproducible data even on difficult samples and/or underrepresented populations. The current configuration of the instrument includes a blue laser (488 nm) and a violet laser (405 nm) for readings of up to 10 colors. The instrument is also equipped with an autosampler with high-throughput plate reading capability (96 and/or 384 wells), ensuring fast and repeatable acquisitions on large amounts of samples

Integra Assist Plus. Instrument capable of performing different pipetting methods with very high accuracy and speed, ensuring operator-dependent error reduction, high reproducibility, and accelerated work on high-throughput experiments. Different configurations for plates, reservoirs and tube housings are possible.

F.A.S.T Formulatrix. Automatic high-throughput dispensing instrument with 96-channel head. With "Positive Displacement" technology, it allows with high precision, the dispensing of small volumes (0.1 µl), without limits related to the Chemistry characteristics of the solutions to be dispensed or aliquoted. Adaptable to plates of different sizes and multifunctional (Cell-Based Assay, Antibody Assay and Enzymatic Assay, nucleic acid extraction and purification).

SPARK Tecan. It offers luminescence, fluorescence intensity, UV absorbance (UV-VIS) and time-resolved fluorescence technologies useful for biochemical and metabolic assays. With the addition of some modules it can easily be upraded to monitor in "real time" imaging of cells in plates from 6 up to 384 well in high-throughput

ChemiDoc Biorad. It is a digital protein band acquisition and visualization system equipped with high sensitivity. It exploits both fluorescence and ECL technologies by increasing sensitivity and the ability to detect multiple targets using fluorophores with non-overlapping excitation and emission spectra

MACSQuant Tyto Cell Sorter Milteny: The MACSQuant_Tyto is an innovative cell sorter based on a single-use, sterile cartridge (MACSQuant Tyto Cartridge). Inside the cartridge, sorting is done by a microchip-controlled valve that directs cells into the collection chamber at low pressure, reducing cell stress and maintaining sample sterility

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Phenocycler-Fusion 2.0 AKOYA: The Phenocycler-Fusion is a spatial proteomics tool that introduces automation, with rapid fluidics, reducing staining and image acquisition times. The antibody barcoding used by the AKOYA/QUANTERIX technology preserves the integrity of the sample, allowing it to be used for transcriptomics analysis as well. In addition, the high-resolution microscope, which can also be used without automation, allows simultaneous acquisition of 6 different colors, in addition to DAPI.

Singulator S2 Genomics: The Singulator is an instrument that allows dissociation of solid tissues into live cells, usable in viable assays, or nuclei usable for genomics and transcriptomics assays such as ATAC-seq. The instrument is easy to use and allows the extraction conditions to be adapted according to the tissue of use by temperature control, addition of digestion enzymes, and modulation of the dissociation rate within sterile disposable cardriges.

Stabularium:

The facility is available at the University Biotechnology Complex on V. Volturno. It is organized in such a way as to allow the handling and suitable maintenance of animals, respecting their needs. The rooms are arranged to provide an environment that takes into account the physiological and ethological needs of the species housed and, in all rooms, there are controlled conditions of lighting, temperature and humidity. The enclosure is also equipped with an environment suitable for the maintenance of immunocompromised mice, with thermostatted enclosure cabinets and appropriate filters for air purification. Use of the enclosure ensures validation of in vitro studies and models, facilitating faster clinical translation.