9 research outputs found
Mechanical thrombectomy in acute stroke – Five years of experience in Poland
Objectives
Mechanical thrombectomy (MT) is not reimbursed by the Polish public health system. We present a description of 5 years of experience with MT in acute stroke in Comprehensive Stroke Centers (CSCs) in Poland.
Methods and results
We retrospectively analyzed the results of a structured questionnaire from 23 out of 25 identified CSCs and 22 data sets that include 61 clinical, radiological and outcome measures.
Results
Most of the CSCs (74%) were founded at University Hospitals and most (65.2%) work round the clock. In 78.3% of them, the working teams are composed of neurologists and neuro-radiologists. All CSCs perform CT and angio-CT before MT. In total 586 patients were subjected to MT and data from 531 of them were analyzed. Mean time laps from stroke onset to groin puncture was 250±99min. 90.3% of the studied patients had MT within 6h from stroke onset; 59.3% of them were treated with IV rt-PA prior to MT; 15.1% had IA rt-PA during MT and 4.7% – emergent stenting of a large vessel. M1 of MCA was occluded in 47.8% of cases. The Solitaire device was used in 53% of cases. Successful recanalization (TICI2b–TICI3) was achieved in 64.6% of cases and 53.4% of patients did not experience hemorrhagic transformation. Clinical improvement on discharge was noticed in 53.7% of cases, futile recanalization – in 30.7%, mRS of 0–2 – in 31.4% and mRS of 6 in 22% of cases.
Conclusion
Our results can help harmonize standards for MT in Poland according to international guidelines
Structural and Functional Characterization of the Hemolymphangiogenic Microenvironment in Lung Cancer
Background: The hypothesis of the existence of a Lung Cancer Initiating Cell (LCIC) offers
new pathogenetic and therapeutic options. CICs express vascular-related molecules in
order to induce neoangiogenesis and establish an aberrant vascular niche. Conversely,
tumor angiogenesis and formation of a cancer vascular niche contribute to the maintenance
of CIC. The limited success of anti-angiogenic strategies in lung cancer imposes a better
knowledge of the biology and architecture of the tumor vascular microenvironment. The
aim of our study was to characterize the structural and functional changes of blood and
lymphatic vasculature in human lung cancer. Methods: Fresh samples of the neoplastic(T) and spared distal (Dist) lung from 30 patients affected by NSCLC (21 Adeno and
9 Squamous) and 9 Neuroendocrine tumors were processed for immunohistochemical
analysis and cell isolation. Control healthy lung (CTRL) was represented by 12 samples
collected at autopsy from patients who died in the absence of respiratory diseases and
6 surgical specimens of pneumothorax. Immunofluorescence and confocal microscopy
were employed using specific antibodies to detect blood (CD31, vWF, a-SMA) and
lymphatic (Podoplanin Pdn, Lyve-1 and Prox-1) vessels. Moreover, the distribution of cells
expressing stem/progenitor cell associated antigens (c-kit, CD133, CD34 and PDGFR)
was assessed. Phenotypical and functional characterization on immunomagnetically
sorted hematic (CD31pos) and lymphatic (CD31pos/Pdnpos) endothelial cells was
performed on cells isolated from Dist and T lung samples. Results: Results indicated
that, compared to CTRL capillary density increased by 77% and 74% in Dist and T
portions of the lung, respectively. Conversely, the numerical incidence of venules did not
show significant difference. These parameters were similarly represented in the three
different tumor histotypes. A significant increase in arteriolar density was observed in
all tumor types compared to CTRL. Moreover, the number of arterioles within T was
increased by 3-fold compared to Dist portion. The quantitative analysis of lymphatic
vessels showed similar values in all types of cancer specimens although rarefaction of
these vascular structures was observed compared to CTRL. Moreover, lymphatic vessels
density was 10-fold higher in Dist lung than in T. Immunofluorescence confocal analysis
documented a positive gradient of c-kitpos, CD34pos and PDGFRpos progenitor cells
from Dist towards T in all cancer samples. However, compared to CTRL, cells expressing
c-kit and CD34 were more numerous both in the Dist and T portion of the lung, while
the increase in PDGFRpos cells was present only in T. Blood (BEC) and lymphatic (LEC)
endothelial cells isolated from Dist and T samples of the lung, showed different growth
properties and variable expression of Tumor Endothelial Marker (TEM) and receptor
tyrosine kinases as VEGFR 2 and 3, PDGFRbeta, EGFR, IGF-1R, and c-met. Functional
assays indicated that T derived LEC possess higher tube forming ability on matrigel
than Dist LEC while this phenomenon was not observed in BEC. Moreover, wound healing
assay showed a VEGF-C independent reduced migratory capacity of LEC isolated
from T samples compared to the Dist counterpart. Conclusion: Specific changes
in the composition and function of the tumor hemolymphangiogenic environment
occur in lung cancer providing innovative pathogenetic and therapeutic approaches
Differential Tumorigenic Properties of Mesenchymal Cells From Neoplastic and Non-Neoplastic Human Lung in NSCLC
the pathogenesis and management of lung cancer. A relevant component of the niche
is represented by supportive stromal cells that control the fate of CICs by a reciprocal
cross-talk. The understanding of these cellular events could represent a significant
advancement in cancer biology and treatment. Recent observations by our and other
laboratories have suggested that mesenchymal stromal cells (MSC) regulate lung
cancer growth and resistance, thus generating large expectations in novel anti-cancer
strategies. The aim of our study was to determine whether MSC isolated from NSCLC
and from non-neoplastic human lung samples possess different biologic properties and
tumorigenic potential. Methods: Fresh samples of neoplastic and spared lungs from 58
male patients (80% smokers) affected by primary pulmonary adenocarcinoma undergoing
surgical resection were processed. Stromal cells were separated from epithelial cells by
negative selection using EpCAM (CD326)-based immunomagnetic sorting. After further
enrichment, we could expand for at least 14 passages a population of CD90, CD105,
CD73 and CD44 positive MSC from lung cancer (Lc-MSC) and non-neoplastic (Nn-MSC)
lung tissue. The oncogenic potential of these cells from the same patient was tested on
a Calu-3-based in vitro model of NSCLC by co-culture and conditioned media (CM) and
in vivo by xenotransplantation in Balb/c Nude mice. In vivo cell tracking was achieved by
pre-labeling MSC with Quantum dots 585 (Qdots). Morphometric assessment of tissue
composition and immunofluorescence combined with FISH analysis of human X and Y
chromosomes was performed on xenografted tumors. Results: Nearly 30x106 cells
could be typically obtained after 3 passages in each case, however, compared to Nn-
MSC, cultures of Lc-MSC displayed lower growth kinetic and mitotic index while higher
survival and HIF-1-alpha (Hypoxia-inducible-factor-1) upregulation in response to hypoxia
was observed. A larger fraction of Lc-MSC expressed transcription factors involved in
stemness (Oct3/4, SOX2) and in bronchioalveolar (TTF1, ETS-1, CCL10) commitment.
Co-cultures demonstrated that Lc-MSC significantly increased Calu-3 growth as
compared to Nn-MSC in transwell assay and by contact. CM from Lc-MSC similarly
promoted Calu-3 expansion as compared to Nn-MSC. When 2.5x106 Lc-MSC or Nn-
MSC from the same patient were subcutaneously co-injected with Calu-3, a 38% and
17% increase in tumor volume was respectively observed, compared to the injection
of an equal number of Calu-3 alone (CTRL). Lc-MSC or Nn-MSC injected alone did not
generate tumors. Quantitative estimation of the in vivo expansion of neoplastic cells
indicated that the addition of Lc-MSC increased by 6-fold and 29-fold Calu-3 replication
compared to Nn-MSC and CTRL, respectively. Cell tracking documented that Qdots
labelled MSC were located at the boundary of neoplastic epithelial glands generated by
X-chromosome polysomic Calu-3 cells. A comparative molecular analysis of Lc-MSC and
Nn-MSC is ongoing for the identification of distinctive signalling pathways implicated in
the microenvironemental control of CIC on NSCLC development. Conclusion: Profound
differences exist in the biology and oncogenic potential of intratumoral and
normal lung MSC strongly supporting the notion that the tumor microenvironment
may represent a potential target of new customized therapeutic strategies
Differential Tumorigenic Properties of Mesenchymal Cells From Neoplastic and Non-Neoplastic Human Lung in NSCLC
the pathogenesis and management of lung cancer. A relevant component of the niche
is represented by supportive stromal cells that control the fate of CICs by a reciprocal
cross-talk. The understanding of these cellular events could represent a significant
advancement in cancer biology and treatment. Recent observations by our and other
laboratories have suggested that mesenchymal stromal cells (MSC) regulate lung
cancer growth and resistance, thus generating large expectations in novel anti-cancer
strategies. The aim of our study was to determine whether MSC isolated from NSCLC
and from non-neoplastic human lung samples possess different biologic properties and
tumorigenic potential. Methods: Fresh samples of neoplastic and spared lungs from 58
male patients (80% smokers) affected by primary pulmonary adenocarcinoma undergoing
surgical resection were processed. Stromal cells were separated from epithelial cells by
negative selection using EpCAM (CD326)-based immunomagnetic sorting. After further
enrichment, we could expand for at least 14 passages a population of CD90, CD105,
CD73 and CD44 positive MSC from lung cancer (Lc-MSC) and non-neoplastic (Nn-MSC)
lung tissue. The oncogenic potential of these cells from the same patient was tested on
a Calu-3-based in vitro model of NSCLC by co-culture and conditioned media (CM) and
in vivo by xenotransplantation in Balb/c Nude mice. In vivo cell tracking was achieved by
pre-labeling MSC with Quantum dots 585 (Qdots). Morphometric assessment of tissue
composition and immunofluorescence combined with FISH analysis of human X and Y
chromosomes was performed on xenografted tumors. Results: Nearly 30x106 cells
could be typically obtained after 3 passages in each case, however, compared to Nn-
MSC, cultures of Lc-MSC displayed lower growth kinetic and mitotic index while higher
survival and HIF-1-alpha (Hypoxia-inducible-factor-1) upregulation in response to hypoxia
was observed. A larger fraction of Lc-MSC expressed transcription factors involved in
stemness (Oct3/4, SOX2) and in bronchioalveolar (TTF1, ETS-1, CCL10) commitment.
Co-cultures demonstrated that Lc-MSC significantly increased Calu-3 growth as
compared to Nn-MSC in transwell assay and by contact. CM from Lc-MSC similarly
promoted Calu-3 expansion as compared to Nn-MSC. When 2.5x106 Lc-MSC or Nn-
MSC from the same patient were subcutaneously co-injected with Calu-3, a 38% and
17% increase in tumor volume was respectively observed, compared to the injection
of an equal number of Calu-3 alone (CTRL). Lc-MSC or Nn-MSC injected alone did not
generate tumors. Quantitative estimation of the in vivo expansion of neoplastic cells
indicated that the addition of Lc-MSC increased by 6-fold and 29-fold Calu-3 replication
compared to Nn-MSC and CTRL, respectively. Cell tracking documented that Qdots
labelled MSC were located at the boundary of neoplastic epithelial glands generated by
X-chromosome polysomic Calu-3 cells. A comparative molecular analysis of Lc-MSC and
Nn-MSC is ongoing for the identification of distinctive signalling pathways implicated in
the microenvironemental control of CIC on NSCLC development. Conclusion: Profound
differences exist in the biology and oncogenic potential of intratumoral and
normal lung MSC strongly supporting the notion that the tumor microenvironment
may represent a potential target of new customized therapeutic strategies
Cardioprotection by Targeting the Pool of Resident and Extracardiac Progenitors
The adult heart has the capacity to generate new myocytes that is markedly enhanced in acute and chronic heart failure of ischemic and non-ischemic origin. In addition, a pool of blood trafficking progenitor cells able to sense myocardial damage may home to the sites of injury participating to cardiac repair. This new view of myocardial biology leads to an expanding long-term research and therapeutic goals for cardioprotection. A fundamental concept to be analyzed is whether cardiac diseases are influenced by changes in the properties of tissue specific and circulating progenitors. Loss of self-renewal capacity, impaired growth or increased susceptibility to death may lead to a reduction of progenitors and leave myocardial damage unrepaired. Cardiac progenitors generate all myocardial cell lineages, thus impairment in their growth is expected to be critically involved in the structural and functional modifications of the heart. The fact that, in addition to well known effects of anthracyclines, also new drugs that target molecular pathways implicated in cell death and growth can be cardiotoxic further supports our hypothesis. Understanding the role of resident and extracardiac progenitors in the pathogenesis of cardiomyopathies of different etiology will provide not only a better comprehension of cardiac homeostasis but will also open new avenues for therapeutic interventions. The progress toward effective myocardial regeneration based on exploiting the self-renewal potential of the myocardium and the systemic pool of cardiogenic cells shouldadvance the likelihood of efficient cardioprotection and restoration of cardiac function
Low PD-1 expression in Cytotoxic CD8+ Tumor infiltrating Lymphocytes Confers an Immune Privileged Tissue Microenvironment in NSCLC with a Prognostic and Predictive Value
The success of immune checkpoint inhibitors strengthens the notion that tumor growth and regression are immune regulated. To determine whether distinct tissue immune microenvironments differentially impact on clinical outcome in Non Small Cell Lung Cancer (NSCLC), an extended analysis of PD-L1 and Tumor Infiltrating Lymphocytes (TILs) was performed. Experimental Design Samples from resected adenocarcinoma (ADC 42) and squamous cell carcinoma (SCC 58) and from 26 advanced diseases (13 ADC, 13 SCC) treated with nivolumab were analyzed. PD-L1 expression and the incidence of CD3, CD8, CD4, PD-1, CD57, FOXP3, CD25 and Granzyme B TILs was immunohistochemically assessed. Results PD-L1 levels inversely correlated with N involvement although did not show a statistical significant prognostic value in resected patients. The incidence and phenotype of TILs differed in SCC vs ADC in which EGFR and KRAS mutations conditioned a different frequency and tissue localization of lymphocytes. NSCLC resected patients with high CD8 pos lymphocytes lacking PD-1 inhibitory receptor had a longer Overall Survival (OS:HR=2.268, 95%CI 1.056-4.871,p=0.03). PD-1-to-CD8 ratio resulted a prognostic factor both on univariate (HR=1.952, 95%CI 1.34-3.12,p=0.001) and multivariate (HR=1.943, 95%CI 1.38-2.86,p=0.009) analysis. Moreover, low PD-1 incidence among CD8(pos) cells was a distinctive feature of nivolumab treated patients showing clinical benefit with a prolonged Progression-Free Survival (PFS:HR=4.51, 95%CI 1.45-13.94,p=0.004). Conclusions In the presence of intrinsic variability in PD-L1 expression, the reservoir of PD-1 negative effector T-lymphocytes provides an immune-privileged microenvironment with a positive impact on survival of patients with resected disease and response to immunotherapy in advanced NSCLC
Mechanical thrombectomy in acute stroke : five years of experience in Poland
Objectives: Mechanical thrombectomy (MT) is not reimbursed by the Polish public health
system. We present a description of 5 years of experience with MT in acute stroke in
Comprehensive Stroke Centers (CSCs) in Poland.
Methods and results: We retrospectively analyzed the results of a structured questionnaire
from 23 out of 25 identified CSCs and 22 data sets that include 61 clinical, radiological and
outcome measures.
Results: Most of the CSCs (74%) were founded at University Hospitals and most (65.2%) work
round the clock. In 78.3% of them, the working teams are composed of neurologists and
neuro-radiologists. All CSCs perform CT and angio-CT before MT. In total 586 patients were
subjected to MT and data from 531 of them were analyzed. Mean time laps from stroke onset
to groin puncture was 250 99 min. 90.3% of the studied patients had MT within 6 h from
stroke onset; 59.3% of them were treated with IV rt-PA prior to MT; 15.1% had IA rt-PA during
MT and 4.7% - emergent stenting of a large vessel. M1 of MCA was occluded in 47.8% of cases.
The Solitaire device was used in 53% of cases. Successful recanalization (TICI2b–TICI3) was
achieved in 64.6% of cases and 53.4% of patients did not experience hemorrhagic transformation. Clinical improvement on discharge was noticed in 53.7% of cases, futile recanalization - in 30.7%, mRS of 0–2 - in 31.4% and mRS of 6 in 22% of cases.
Conclusion: Our results can help harmonize standards for MT in Poland according to international guideline