Periodic Reporting for period 3 - XHaLe (Hanover experimental lung research project)
Reporting period: 2021-06-01 to 2022-11-30
Assigning diseases to a location within the body and drawing conclusions about etiology and pathogenesis from anatomical changes make up the traditional core method of pathology, described already by Rudolph Virchow as „the anatomical concept of pathology“.
Non-neoplastic lung diseases (NNLD), such as COPD and interstitial lung diseases rank second among the causes of death and NNLD appear as a growing challenge for Germany, the EU and worldwide. Viruses, are assumed as exacerbating co-factors of fibrotic lung diseases. These are characterized by irreversible fibrotic remodelling with a relentless loss of lung function and a 5-year survival of only 30%. Pulmonary fibrosis results from aberrant injury-repair responses to chronic lung injury. Over the last 2,5 years, in Xhale, my team and I were dedicated to unravel the mechanisms of pathogenesis by looking at the cellular and molecular changes that are found in human lung explants and correlate them with morphological and clinical aspects of these diseases. Using these molecular Patterns we were able to better define these diseases and set them apart from each other.
Of special interest in this regard is the question of the origin of these NNLDs. Here, we found recently evidence for initiation of fibrotic remodelling by SARS-CoV-2 in humans (Ackermann, et al., 2020, NEJM) on the level of newly formed blood vessels, in a pattern associated with pulmonary fibrosis (Jonigk et al., 2020). Although the precise temporal sequence of events and mechanisms of disease are not understood, some characteristic pathobiological characteristics are recognized.
Non-neoplastic lung diseases (NNLD), such as COPD and interstitial lung diseases rank second among the causes of death and NNLD appear as a growing challenge for Germany, the EU and worldwide. Viruses, are assumed as exacerbating co-factors of fibrotic lung diseases. These are characterized by irreversible fibrotic remodelling with a relentless loss of lung function and a 5-year survival of only 30%. Pulmonary fibrosis results from aberrant injury-repair responses to chronic lung injury. Over the last 2,5 years, in Xhale, my team and I were dedicated to unravel the mechanisms of pathogenesis by looking at the cellular and molecular changes that are found in human lung explants and correlate them with morphological and clinical aspects of these diseases. Using these molecular Patterns we were able to better define these diseases and set them apart from each other.
Of special interest in this regard is the question of the origin of these NNLDs. Here, we found recently evidence for initiation of fibrotic remodelling by SARS-CoV-2 in humans (Ackermann, et al., 2020, NEJM) on the level of newly formed blood vessels, in a pattern associated with pulmonary fibrosis (Jonigk et al., 2020). Although the precise temporal sequence of events and mechanisms of disease are not understood, some characteristic pathobiological characteristics are recognized.
Our study so far provided important insights in the molecular background of specific NNLDs, their possible onset, the involvement of virusses and the molecular characteristics.
The detals of our studies can be found here:
Verleden SE and Jonigk v 3d Virtual Pathohistology of Lung Tissue from Covid-19 Patients based on Phase Contrast X-ray Tomography. eLife 2020 Aug 20. doi: 10.7554/elife.60408. PMID:32815517 (Impact Factor 2019: 6.830)
*contributed equally
Ackermann M, et al. and Jonigk Pulmonary Vascular Endothelialitis, Thrombosis and Angiogenesis in COVID-19. N Engl J Med. 2020 May 21. doi: 10.1056/NEJMoa2015432. PMID: 32437596 (Impact- Factor 2019: 70.670)
Ackermann M, et al. and Jonigk Pulmonary Vascular Pathology in Covid-19. Reply. N Engl J Med. 2020 Jul 17. doi: 10.1056/NEJMc2022068. PMID: 32678534. (Impact Factor 2019: 70.670)
Neubert L*, at al and Jonigk D#. Molecular Profiling of Vascular Remodeling in Chronic Pulmonary Disease. Am J Pathol. 2020 Apr 7. pii: S0002-9440(20)30145-0. doi: 10.1016/j.ajpath.2020.03.008 (Impact Factor 2017: 4.189)
Ackermann M, et al and Jonigk D. Morphomolecular motifs of pulmonary neoangiogenesis in interstitial lung diseases. Eur Respir J. 2020 Mar 12;55(3):1900933. doi: 10.1183/13993003.00933-2019 PMID: 31806721. (Impact Factor 2018: 12,242)
Jonigk D, et al. Morphological and molecular motifs of fibrosing pulmonary injury patterns. J Pathol Clin Res. 2019 doi: 10.1002/cjp2.141. (Impact Factor 2015: 7.381)
Neubert L* at al and Jonigk D°. Comprehensive Three-dimensional Morphology of Neoangiogenesis in Pulmonary Veno-occlusive Disease and Pulmonary Capillary Hemangiomatosis. J Pathol Clin Res 2019; 5(2): 108-114. doi: 10.1002/cjp2.125. (Impact Factor 2015: 7.381)
Ackermann M et al. A 3D iPSC-differentiation model identifies interleukin-3 as a regulator of early human hematopoietic specification. Haematologica 2020 Apr 23. pii: haematol.2019.228064. doi: 10.3324/haematol.2019.228064 (Impact Factor 2018: 7.570)
Ackermann M, et al. Bioreactor-based mass production of human iPSC-derived macrophages enables immunotherapies against bacterial airway infections. Nat Commun 2018; 9(1): 5088. doi: 10.1038/s41467-018-07570-7. (Impact Factor 2017: 13.691)
WP 1: "Comprehensive and standardized work-up of explanted human lungs"
This is an ongoing endevour and the backbone of our work. So far we were able to include 99% of all lungs transplanted at Hannover Medical School into the Xhale project (113 lungs in 2018, 104 lungs in 2019 and 88 lungs in 2020)
WP 2: "analysis of human lung explants - characterization of the inflammome in NNLDs"
This WP is now a standardized part of our routine workup of lungs and allowed us great insights into the pathobiology of differend NNLDs. We were able to set apart different NNLDs due to their underlying molecular gene expression characteristics and provide predictive algorithmy that are able to identify a fibrotic pattern with *90% certainty".
WP 3: From predicted pathomechanisms to certainty - analysis of precision-cut lung slices (PCLS) from explants
The workpackage under construction. The tissue culture of human lung tissue is set up and described up to 6 weeks of culture in detail regarding viability of different cell types, gene expression and histology (Manuscripts under development) .
Macrophage subtypes of various kinds are now available via standardized culture and macrophages cultures containing specific gene modifications theat derived from the analysis of WP 2 are being constructed currently. This work will be a main focus of the second half of Xhale.
WP 4: Molecular investigation of the angiogeneic driver of NNLD remodelling
We were also able to describe and identify specific vascular changes specifically associated with different forms of fibrotic NNLD remodelling. Of special note here is the involvement of intussusceptive neoangiogenesis, which is not only a typical trait of Alveolar fibroelastosis and NSIP, but also of COVID-19 lung pathology, that might be involved with the long term complications that SARS-CoV-2 survivors suffer from and it also presents a possible molecular access point for inhibitory therapy.
The detals of our studies can be found here:
Verleden SE and Jonigk v 3d Virtual Pathohistology of Lung Tissue from Covid-19 Patients based on Phase Contrast X-ray Tomography. eLife 2020 Aug 20. doi: 10.7554/elife.60408. PMID:32815517 (Impact Factor 2019: 6.830)
*contributed equally
Ackermann M, et al. and Jonigk Pulmonary Vascular Endothelialitis, Thrombosis and Angiogenesis in COVID-19. N Engl J Med. 2020 May 21. doi: 10.1056/NEJMoa2015432. PMID: 32437596 (Impact- Factor 2019: 70.670)
Ackermann M, et al. and Jonigk Pulmonary Vascular Pathology in Covid-19. Reply. N Engl J Med. 2020 Jul 17. doi: 10.1056/NEJMc2022068. PMID: 32678534. (Impact Factor 2019: 70.670)
Neubert L*, at al and Jonigk D#. Molecular Profiling of Vascular Remodeling in Chronic Pulmonary Disease. Am J Pathol. 2020 Apr 7. pii: S0002-9440(20)30145-0. doi: 10.1016/j.ajpath.2020.03.008 (Impact Factor 2017: 4.189)
Ackermann M, et al and Jonigk D. Morphomolecular motifs of pulmonary neoangiogenesis in interstitial lung diseases. Eur Respir J. 2020 Mar 12;55(3):1900933. doi: 10.1183/13993003.00933-2019 PMID: 31806721. (Impact Factor 2018: 12,242)
Jonigk D, et al. Morphological and molecular motifs of fibrosing pulmonary injury patterns. J Pathol Clin Res. 2019 doi: 10.1002/cjp2.141. (Impact Factor 2015: 7.381)
Neubert L* at al and Jonigk D°. Comprehensive Three-dimensional Morphology of Neoangiogenesis in Pulmonary Veno-occlusive Disease and Pulmonary Capillary Hemangiomatosis. J Pathol Clin Res 2019; 5(2): 108-114. doi: 10.1002/cjp2.125. (Impact Factor 2015: 7.381)
Ackermann M et al. A 3D iPSC-differentiation model identifies interleukin-3 as a regulator of early human hematopoietic specification. Haematologica 2020 Apr 23. pii: haematol.2019.228064. doi: 10.3324/haematol.2019.228064 (Impact Factor 2018: 7.570)
Ackermann M, et al. Bioreactor-based mass production of human iPSC-derived macrophages enables immunotherapies against bacterial airway infections. Nat Commun 2018; 9(1): 5088. doi: 10.1038/s41467-018-07570-7. (Impact Factor 2017: 13.691)
WP 1: "Comprehensive and standardized work-up of explanted human lungs"
This is an ongoing endevour and the backbone of our work. So far we were able to include 99% of all lungs transplanted at Hannover Medical School into the Xhale project (113 lungs in 2018, 104 lungs in 2019 and 88 lungs in 2020)
WP 2: "analysis of human lung explants - characterization of the inflammome in NNLDs"
This WP is now a standardized part of our routine workup of lungs and allowed us great insights into the pathobiology of differend NNLDs. We were able to set apart different NNLDs due to their underlying molecular gene expression characteristics and provide predictive algorithmy that are able to identify a fibrotic pattern with *90% certainty".
WP 3: From predicted pathomechanisms to certainty - analysis of precision-cut lung slices (PCLS) from explants
The workpackage under construction. The tissue culture of human lung tissue is set up and described up to 6 weeks of culture in detail regarding viability of different cell types, gene expression and histology (Manuscripts under development) .
Macrophage subtypes of various kinds are now available via standardized culture and macrophages cultures containing specific gene modifications theat derived from the analysis of WP 2 are being constructed currently. This work will be a main focus of the second half of Xhale.
WP 4: Molecular investigation of the angiogeneic driver of NNLD remodelling
We were also able to describe and identify specific vascular changes specifically associated with different forms of fibrotic NNLD remodelling. Of special note here is the involvement of intussusceptive neoangiogenesis, which is not only a typical trait of Alveolar fibroelastosis and NSIP, but also of COVID-19 lung pathology, that might be involved with the long term complications that SARS-CoV-2 survivors suffer from and it also presents a possible molecular access point for inhibitory therapy.
The most striking progress beyond the state of the art was our newly formed cooperation with Professor Peter Lee and colleagues. Together we performed 3D invetigations of not only SARS-CoV-2 infected lungs at the ESRF in Grenoble, but also from approximatly 200 lung samples from our WP1 collected lungs. The result of these investigations will continue to gain insights over the next 5-10 yeaars since we are now able to see whole human lungs in cellular resolution allowing for much deeper analysis of the remodelled architecture than current stae of the art radiological approaches.
At the same time our cooperation with Professor Tim Saldit from Göttingen allowed us to analyze human lung samples embedded in paraffin in 3D using X-ray tomography and decribe the 3D continuum of hyalin membranes in COVID-19 lung samples, one of the defining features of COVID-19 lungs and a major reason why SARS-CoV-2 infected lungs fail to provide sufficient oxygen for the patients.
All these efforts will provide much deeper insights in pulmonary diseases and be involved in therapeutic solutions.
At the same time our cooperation with Professor Tim Saldit from Göttingen allowed us to analyze human lung samples embedded in paraffin in 3D using X-ray tomography and decribe the 3D continuum of hyalin membranes in COVID-19 lung samples, one of the defining features of COVID-19 lungs and a major reason why SARS-CoV-2 infected lungs fail to provide sufficient oxygen for the patients.
All these efforts will provide much deeper insights in pulmonary diseases and be involved in therapeutic solutions.