Stem Cells Heal Damaged Cells by Transferring Mitochondria

An Indian team from Delhi, India has identified a protein that increases the transfer of mitochondria from mesenchymal stem cells to lung cells, thus augmenting the healing of lung cells.

Stem cells like mesenchymal stem cells from bone marrow, fat, tendons, liver, skeletal muscle, and so on secrete a host of healing molecules, but they also form bridges to other cells and export their own mitochondria to heal damaged cells. Mitochondria are the structures inside cells that make energy. Damaged cells can have serious energy deficiencies and mitochondrial transfer ameliorates such problems (see Cárdenes N et al, Respiration. 2013;85(4):267-78).

This present work from the laboratory of Anurag Agrawal, who is housed in the Centre of Excellence in Asthma & Lung Disease, at the CSIR‐Institute of Genomics and Integrative Biology in Delhi, India has identified a protein called Miro1 that regulates the transfer of mitochondria to recipient cells.

Mitochondrial transfer has so many distinct benefits that stem cell scientists hope to engineer stem cells to transfer more of their mitochondria to damaged cells, and Miro1 might be a target for such stem cell engineering experiments.

Mitochondrial transfer between stem cells and other cells occurs by means of tunneling nanotubes, which are thread-like structures formed from the plasma membranes of cells that form bridges between different cell types. Under stressful conditions, the number of these nanotubes increases.

In the present study. stem cells engineered to express more Miro1 protein transferred mitochondria more efficiently than control stem cells. When used in mice with damaged lungs and airways, these Miro1-overexpressing cells were therapeutically more effective than control cells.

This study presents the first mechanistic insight into how Mesenchymal Stem Cells (MSC) act as mitochondrial donors during attenuation of lung inflammation and injury. Mitochondrial donation is an essential part of the MSC therapeutic effect in these models and is positively regulated by Miro1 / Rhot1 mitochondrial transport proteins.
This study presents the first mechanistic insight into how Mesenchymal Stem Cells (MSC) act as mitochondrial donors during attenuation of lung inflammation and injury. Mitochondrial donation is an essential part of the MSC therapeutic effect in these models and is positively regulated by Miro1 / Rhot1 mitochondrial transport proteins.

The hope is to use Miro1 manipulations to make better stem cell therapies for human diseases.

To summarize this work:

1. MSCs donate mitochondria to stressed epithelial cells (EC) that have malfunctioning mitochondrial.  Cytoplasmic nanotubular bridges form between the cells and Miro‐1 mediated mitochondrial transfer occurs unidirectionally from MSCs to ECs.

2. Other mesenchymal cells like smooth muscle cells and fibroblasts express Miro1 and can also donate mitochondria to ECs, but with low efficiency. ECs have very low levels of Miro1 and, as a rule, do not donate mitochondria.

3. Enhanced expression of Miro1 in mesenchymal cells increases their mitochondrial donor efficiency.  Conversely, cells lacking Miro1 do not show MSC mediated mitochondrial donation.

4. Miro1‐overexpressing MSCs have enhanced therapeutic effects in three different models of allergic lung inflammation and rat poison-induced lung injury.  Conversely, Miro1‐depleted MSCs lose much of their therapeutic effect.  Miro1 overexpression in MSCs may lead to more effective stem cell therapy.

Culture Media from Mesenchymal Stem Cells Heals Injured Lungs

Acute lung injury and acute respiratory distress syndrome remain major causes of death and suffering despite advances in management of these conditions. The incidence of these conditions is expected to double in the next 25 years, and treatment for it is largely supportive.

Fortunately, mesenchymal stem cells (MSCs) from bone marrow have been used in experimental models to treat lung injury in rodents. MSCs can engraft into lung tissue and become lung tissue (or at least turn into cells that sure look a whole lot like lung tissue). MSCs can also suppress the types of immune responses that tend to really chew up lung tissue. Thus, MSC administration seems to improve the condition of lungs that have been attacked by infections or damaging agents.

However, the rates at which MSCs engraft into lung tissue is rather low; too low, in fact, to account for the benefit provided by MSCs. Therefore, MSCs appear to help repair lung tissue by means of “paracrine” mechanisms. This 50-cent word simply means that MSCs repair the lung by secreting molecules that promote lung healing.

To test this hypothesis, researchers in the laboratory of Bernard Thérband from the Ottawa Hospital Research Institute in Ottawa, Canada has grown MSCs in culture, and used the growth medium after the MSCs had been removed from it to treat mice that suffered from lung injuries.

To induce lung injury, mice were treated with isolated bits of bacterial cells that are known to promote acute lung injury. Then a group of these lung-injured mice were treated with conditioned medium from bone marrow MSCs that had been grown in culture dishes.

The MSC-conditioned medium decreased lung inflammation, and disruptions of the blood vessels in the lung normally observed during lung injury. Therefore, the lungs did not fill up with liquid and pus. However, the conditioned medium did not prevent the weight loss associated with lung injury. The overall tissue architecture of the lung tissue was much more normal in the mice treated with the conditioned medium from MSCs than in the untreated mice. Conditioned medium from other cultured cells had no such sanative effect.

MSC conditioned culture media also modified the activity of white blood cells in the lung. Instead of charging forward into lung tissue and damaging it in response to damage, the white blood cells (so-called “alveolar macrophages”) worked with the lung tissue to help heal it.

Finally, when Thébaud and his colleagues examined the molecules secreted into the medium by the MSCs, they discovered that the culture medium was filled with lots of interesting molecules, but one in particular caught their eye:  Insulin-like growth factor-1 (IGF-1). This molecule has all kinds of healing properties, and it seemed to Thébaud and company that IGF-1 could be responsible for a good portion of the healing. Therefore, they infused the lung-injured mice with purified IGF-1, and, wouldn’t you know, the lungs showed rather robust healing after being damaged with bacterial bits.

Thus MSCs provide lung healing properties and they do so by means of the molecules they secrete. Many of these healing properties can be recapitulated by infusing IGF-1.

Such experiments provide hope that future clinical trials for such treatments are not far off.

Mesenchymal stem cells used to treat Acute Respiratory Distress

Acute Respiratory Distress Syndrome (ARDS) describes a spectrum of increasingly severe acute respiratory failure events. ARDS results from multiple causes that include infections, trauma and major surgery. Clinically, ARDS is the leading cause of death and disability in the critically ill.

The characteristics of ARDS includes a somewhat sudden onset, severe oxygen depletion or hypoxia, stiff lungs that do not expand or contract properly, and the presence of an inflammation in the lungs that results in pulmonary swelling (edema; see Ware LB, Matthay MA. N Engl J Med. 2000, 342:1334–49.).  In the US, there are 200,000 new cases each year, and carries a mortality rate of 40%. This is a mortality rate that is comparable to that seen from HIV infections and breast cancer. The prognosis of ARDS survivors is also somewhat poor. ARDS sufferers can also find themselves fighting with cognitive impairment, depression and muscle weakness. Also ARDS can saddle patients with substantial financial burdens (see Herridge MS, et al. N Engl J Med. 2003, 348:683–93 & Hopkins RO, et al. Am J Respir Crit Care Med. 2005, 171:340–7).

Despite decades of research on ARDS, there are no therapies for it and management of the disease remains supportive.  But now stem cells called “mesenchymal stem cells” offer a potentially successful treatment of ARDS.  Mesenchymal stem cells (MSCs) are multipotent cells stem cells that are derived from adult tissues and capable of self-renewal and can differentiate into cartilage-making cells (chondrocytes), bone-making cells (osteocytes), and fat cells (adipocytes).  Friedenstein and colleagues were the first to isolate MSCs from rodent bone marrow in 1976 ()m the bone marrow in 1976 (see Friedenstein AJ, Gorskaja JF, Kulagina NN. Exp Hematol. 1976, 4:267–74).   Since  their discovery, MSCs have been isolated from many other tissues, including fat, muscle, dermis, placenta, umbilical cord, peripheral blood, liver, spleen, and lung.  The fact that MSCs come from adult tissue, are relatively easy to isolate, and are capable of robust growth in culture, males them attractive candidates for regenerative medicine (see Prockop DJ, et al. J Cell Mol Med. 2010, 14:2190–9).  Additionally, MSCs are usually tolerated by the immune system, which means that they can be transplanted from one individual to another.

Earlier studies provided data that suggested that MSCs actually might differentiate into lung epithelial cells and directly replace the damaged and destroyed lung cells. For example, Kotton et al. demonstrated that bone marrow-derived cells could engraft into pulmonary epithelia and acquire the specific characteristics typical to lung epithelial cells (Kotton DN, et al. Development. 2001, 128:5181–8).  Krause and colleagues showed that transplantation of a single bone marrow-derived blood-cell making (hematopoietic) stem cell could give rise to cells of different organs, including the lung, and demonstrated that up to 20% of lung alveolar cells were derived from this single bone marrow stem cell (Krause DS, et al. Cell. 2001, 105:369–77).  Finally, Suratt and co-workers examined female patients who had received bone marrow transplants from male donors, and found that significant numbers of male bone marrow stem cells, which were detected by the presence of the Y chromosome, had formed cells that engrafted in the lungs of the female patients (Suratt BT, et al. Am J Respir Crit Care Med. 2003, 168:318–2).  Unfortunately, more recent studies have clearly demonstrated that even though MSCs definitely reduce experimental lung injury, engraftment rates are low (see Mei SH, et al. PLoS Med. 2007, 4:e269; & Ortiz LA, et al. Proc Natl Acad Sci U S A. 2007, 104:11002–7). This suggests that direct engraftment of mesenchymal stem cells in the lung is unlikely to be of large therapeutic significance.

Several experiments have suggested many different mechanisms by which MSCs might help injured lungs.  First, MSCs seem to slow down the immune response to lung injury (see Gupta N, et al. J Immunol. 2007, 179:1855–63 & Mei SH, et al. Am J Respir Crit Care Med. 2010, 182:1047–57).  However, instead of acting like classic “anti-inflammatory” drugs might work, MSCs actually decrease host damage that arises from the inflammatory response, but also enhance host resistance to bacterial infections (sepsis).  MSCs decrease the expression of small molecules called “cytokines” that encourage inflammation (see Danchuk S, et al. Stem Cell Res Ther. 2011, 2).  Conversely, they also produce a host of anti-inflammatory molecule (e.g., interleukin 1 receptor antagonist, interleukin-10, and prostaglandin E2; see Németh K, et al. Nat Med. 2009, 15:42–9).  Because of these activities, MSCs reduced the recruitment of white blood cells to the lung during episodes of lung damage.  This is important because when white blood cells are recruited to a damaged area, they act as though they are ticked off and damaged not just the invading bacteria, but anything that stands in their and that includes innocent bystanders.  Thus by keeping ticked off white bloods away from lung tissue, the lung is spared extensive damage.

Secondly, MSCs seem to increase the immune response to sepsis, and reduce lung-damage-induced systemic sepsis.  Sepsis refers to the colonization of the bloodstream by infecting microorganisms.  Damage to the lung epithelium and provide a door from the air we breathe and the bacteria that contaminate it to our bloodstream.  MSCs mitigate lung damage, and therefore, reduce lung-induced sepsis,  MSCs secrete prostaglandin-E2, and this molecule stimulates resident white blood cells in the lung, known as “alveolar macrophages” to produce a molecule called “IL-10.”  IL-10 prevents potentially damaging activated white blood cells from being summoned to the lung (see Németh K, et al. Nat Med. 2009, 15:42–9).   Additionally, MSCs secrete anti-microbial peptides such as LL-37 and  tumour-necrosis-factor-alpha-induced-protein-6 that retard bacterial growth (Krasnodembskaya A, et al. Stem Cells. 2010, 28:2229–3).  When given to mice with lung damaged-induced sepsis, transplanted MSCs increased clearance of bacteria from the lung anf enhanced destruction of the bacteria by resident white blood cells (Mei SH,et al. Am J Respir Crit Care Med. 2010, 182:1047–57).

Thirdly, MSCs aid lung regeneration following injury.  They do this by secreting molecules that protect cells and promote cell survival (so-called “cytoprotective agents”).  MSCs also secrete “angiopoeitin” and “keratinocyte growth factor,” which restore the growth and health of the lung alveolar epithelial and endothelial permeability.  These molecules enhance lung healing in ARDS animals (see Lee JW,et al. Proc Natl Acad Sci U S A. 2009, 106:16357–6Mei SH, et al. PLoS Med. 2007, 4:e269 & Fang X, et al. J Biol Chem 2010. 285:26211–2). 

Clearly MSCs show a very diverse cadre of mechanisms that favorably modulate the immune response, which reduces inflammation and inflammation injury, without compromising the integrity of the immune response.  They also hasten healing of damaged lung tissue.  These features make MSCs attractive therapeutic candidates for ARDS.

Preclinical have proven extremely hopeful.  Human trials are currently in the planning and early stages.  It is not clear what the right dosages of MSCs might be or what is the best way to administer them (intravenous, intra-tracheal, or intra-peritoneal).  Another hurdle is that MSCs are a very heterogeneous population once they are isolated.  Which cells in this mixed population are them best for helping ARDS patients?  All these questions much be addressed before human trials can definitively test MSC treatments for ARDS.